Takahito Watanabe, Sumihare Noji and Taro Mito : GeneKnockout by Targeted Mutagenesis in a Hemimetabolous Insect, the Two-Spotted Cricket Gryllus bimaculatus, using TALENs. In TALENs: Methods and Protocols (Ralf Kuhn et al. eds.), Springer, New York, 2016.
Taro Mito, Yoshiko Inoue, S Kimura, Katsuyuki Miyawaki, Nao Niwa, Yohei Sinmyo, Hideyo Ohuchi and Sumihare Noji : Formation of new organizing regions by cooperation of hedgehog, wingless, and dpp in regeneration of the insect leg; a verification of the boundary model In Morphogenesis and pattern formation in biological systems (Sekimura et al. eds.), Springer, 2003.
T Nohno, Sumihare Noji, E Koyama, F Myokai, Hideyo Ohuchi, K Nishikawa, S Sumitomo, S Taniguchi and Sumihare Noji : Expression patterns of the activin receptor IIA and IIB genes during chick limb development., Wiley-Liss, Inc., New York, 1993.
25.
Sumihare Noji, E Koyama, F Myokai, T Nohno, Hideyo Ohuchi, K Nishikawa and S Taniguchi : Differential expression of three chick FGF receptor genes, FGFR1, FGFR2 and FGFR3, in limb and feather development, Wiley-Liss, Inc., New York, 1993.
Academic Paper (Judged Full Paper):
1.
Takahisa Yamashita, Takahiro Ohde, Taro Nakamura, Yoshiyasu Ishimaru, Takahito Watanabe, Sayuri Tomonari, Yuki Nakamura, Sumihare Noji and Taro Mito : Involvement of the scalloped gene in morphogenesis of the wing margin via regulating cell growth in a hemimetabolous insect Gryllus bimaculatus., Development Growth & Differentiation, Vol.65, No.6, 348-359, 2023.
(Summary)
The acquisition of wings was a key event in insect evolution. As hemimetabolous insects were the first group to acquire functional wings, establishing the mechanisms of wing formation in this group could provide useful insights into their evolution. In this study, we aimed to elucidate the expression and function of the gene scalloped (sd), which is involved in wing formation in Drosophila melanogaster, and in Gryllus bimaculatus mainly during postembryonic development. Expression analysis showed that sd is expressed in the tergal edge, legs, antennae, labrum, and cerci during embryogenesis and in the distal margin of the wing pads from at least the sixth instar in the mid to late stages. Because sd knockout caused early lethality, nymphal RNA interference experiments were performed. Malformations were observed in the wings, ovipositor, and antennae. By analyzing the effects on wing morphology, it was revealed that sd is mainly involved in the formation of the margin, possibly through the regulation of cell proliferation. In conclusion, sd might regulate the local growth of wing pads and influence wing margin morphology in Gryllus.
Shintaro Inoue, Takahito Watanabe, Taiki Hamaguchi, Yoshiyasu Ishimaru, Katsuyuki Miyawaki, Takeshi Nikawa, Akira Takahashi, Sumihare Noji and Taro Mito : Combinatorial expression of ebony and tan generates body color variation from nymph through adult stages in the cricket, Gryllus bimaculatus., PLoS ONE, Vol.18, No.5, 2023.
(Summary)
Insect body colors and patterns change markedly during development in some species as they adapt to their surroundings. The contribution of melanin and sclerotin pigments, both of which are synthesized from dopamine, to cuticle tanning has been well studied. Nevertheless, little is known about how insects alter their body color patterns. To investigate this mechanism, the cricket Gryllus bimaculatus, whose body color patterns change during postembryonic development, was used as a model in this study. We focused on the ebony and tan genes, which encode enzymes that catalyze the synthesis and degradation, respectively, of the precursor of yellow sclerotin N-β-alanyl dopamine (NBAD). Expression of the G. bimaculatus (Gb) ebony and tan transcripts tended to be elevated just after hatching and the molting period. We found that dynamic alterations in the combined expression levels of Gb'ebony and Gb'tan correlated with the body color transition from the nymphal stages to the adult. The body color of Gb'ebony knockout mutants generated by CRISPR/Cas9 systemically darkened. Meanwhile, Gb'tan knockout mutants displayed a yellow color in certain areas and stages. The phenotypes of the Gb'ebony and Gb'tan mutants probably result from an over-production of melanin and yellow sclerotin NBAD, respectively. Overall, stage-specific body color patterns in the postembryonic stages of the cricket are governed by the combinatorial expression of Gb'ebony and Gb'tan. Our findings provide insights into the mechanism by which insects evolve adaptive body coloration at each developmental stage.
Yuki Nakamura, Sayuri Tomonari, Kohei Kawamoto, Takahisa Yamashita, Takahito Watanabe, Yoshiyasu Ishimaru, Sumihare Noji and Taro Mito : Evolutionarily conserved function of the even-skipped ortholog in insects revealed by gene knock-out analyses in Gryllus bimaculatus., Developmental Biology, Vol.485, 1-8, 2022.
(Summary)
Comparing the developmental mechanisms of segmentation among insects with different modes of embryogenesis provides insights on how the function of segmentation genes evolved. Functional analysis of eve by genetic mutants shows that the Drosophila pair-rule gene, even-skipped (eve), contributes to initial segmental patterning. However, eve orthologs tends to have diverse functions in other insects. To compare the evolutionary functional divergence of this gene, we evaluated eve function in a phylogenetically basal insect, the cricket Gryllus bimaculatus. To investigate the phenotypic effects of eve gene knock-out, we generated CRISPR/Cas9 system-mediated mutant strains of the cricket. CRISPR/Cas9 mutagenesis of multiple independent sites in the eve coding region revealed that eve null mutant embryos were defective in forming the gnathal, thoracic, and abdominal segments, consequently shortening the anterior-posterior axis. In contrast, the structures of the anterior and posterior ends (e.g., antenna, labrum, and cercus) formed normally. Hox gene expression in the gnathal, thoracic, and abdominal segments was detected in the mutant embryos. Overall, this study showed that Gryllus eve plays an important role in embryonic elongation and the formation of segmental boundaries in the gnathal to abdominal region of crickets. In the light of studies on other species, the eve function shown in Gryllus might be ancestral in insects.
Tetsuya Bando, Misa Okumura, Yuki Bando, Marou Hagiwara, Yoshimasa Hamada, Yoshiyasu Ishimaru, Taro Mito, Eri Kawaguchi, Takeshi Inoue, Kiyokazu Agata, Sumihare Noji and Hideyo Ohuchi : Toll signalling promotes blastema cell proliferation during cricket leg regeneration via insect macrophages., Development, Vol.149, No.8, 2022.
(Summary)
Hemimetabolous insects, such as the two-spotted cricket Gryllus bimaculatus, can recover lost tissues, in contrast to the limited regenerative abilities of human tissues. Following cricket leg amputation, the wound surface is covered by the wound epidermis, and plasmatocytes, which are insect macrophages, accumulate in the wound region. Here, we studied the function of Toll-related molecules identified by comparative RNA sequencing during leg regeneration. Of the 11 Toll genes in the Gryllus genome, expression of Toll2-1, Toll2-2 and Toll2-5 was upregulated during regeneration. RNA interference (RNAi) of Toll, Toll2-1, Toll2-2, Toll2-3 or Toll2-4 produced regeneration defects in more than 50% of crickets. RNAi of Toll2-2 led to a decrease in the ratio of S- and M-phase cells, reduced expression of JAK/STAT signalling genes, and reduced accumulation of plasmatocytes in the blastema. Depletion of plasmatocytes in crickets using clodronate also produced regeneration defects, as well as fewer proliferating cells in the regenerating legs. Plasmatocyte depletion also downregulated the expression of Toll and JAK/STAT signalling genes in the regenerating legs. These results suggest that Spz-Toll-related signalling in plasmatocytes promotes leg regeneration through blastema cell proliferation by regulating the Upd-JAK/STAT signalling pathway.
Guillem Ylla, Taro Nakamura, Takehiko Itoh, Rei Kajitani, Atsushi Toyoda, Sayuri Tomonari, Tetsuya Bando, Yoshiyasu Ishimaru, Takahito Watanabe, Masao Fuketa, Yuji Matsuoka, A Austen Barnett, Sumihare Noji, Taro Mito and G Cassandra Extavour : Insights into the genomic evolution of insects from cricket genomes., Communications Biology, Vol.4, No.1, 2021.
(Summary)
Most of our knowledge of insect genomes comes from Holometabolous species, which undergo complete metamorphosis and have genomes typically under 2 Gb with little signs of DNA methylation. In contrast, Hemimetabolous insects undergo the presumed ancestral process of incomplete metamorphosis, and have larger genomes with high levels of DNA methylation. Hemimetabolous species from the Orthopteran order (grasshoppers and crickets) have some of the largest known insect genomes. What drives the evolution of these unusual insect genome sizes, remains unknown. Here we report the sequencing, assembly and annotation of the 1.66-Gb genome of the Mediterranean field cricket Gryllus bimaculatus, and the annotation of the 1.60-Gb genome of the Hawaiian cricket Laupala kohalensis. We compare these two cricket genomes with those of 14 additional insects and find evidence that hemimetabolous genomes expanded due to transposable element activity. Based on the ratio of observed to expected CpG sites, we find higher conservation and stronger purifying selection of methylated genes than non-methylated genes. Finally, our analysis suggests an expansion of the pickpocket class V gene family in crickets, which we speculate might play a role in the evolution of cricket courtship, including their characteristic chirping.
(Keyword)
Animals / DNA Methylation / DNA Transposable Elements / Evolution, Molecular / Female / Genes, Insect / Genome, Insect / Gryllidae / Insecta / Male / Phylogeny / Repetitive Sequences, Nucleic Acid / Sequence Analysis, DNA
Yoshiyasu Ishimaru, Sayuri Tomonari, Takahito Watanabe, Sumihare Noji and Taro Mito : Regulatory mechanisms underlying the specification of the pupal-homologous stage in a hemimetabolous insect, Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences, Vol.374, No.1783, 20190225, 2019.
(Summary)
Juvenile hormones and the genetic interaction between the transcription factors Krüppel homologue 1 (Kr-h1) and Broad (Br) regulate the transformation of insects from immature to adult forms in both types of metamorphosis (holometaboly with a pupal stage versus hemimetaboly with no pupal stage); however, knowledge about the exact instar in which this occurs is limited. Using the hemimetabolous cricket Gryllus bimaculatus (Gb), we demonstrate that a genetic interaction occurs among Gb'Kr-h1, Gb'Br and the adult-specifier transcription factor Gb'E93 from the sixth to final (eighth) nymphal instar. Gb'Kr-h1 and Gb'Br mRNAs were strongly expressed in the abdominal tissues of sixth instar nymphs, with precocious adult moults being induced by Gb'Kr-h1 or Gb'Br knockdown in the sixth instar. The depletion of Gb'Kr-h1 or Gb'Br upregulates Gb'E93 in the sixth instar. By contrast, Gb'E93 knockdown at the sixth instar prevents nymphs transitioning to adults, instead producing supernumerary nymphs. Gb'E93 also represses Gb'Kr-h1 and Gb'Br expression in the penultimate nymphal instar, demonstrating its important role in adult differentiation. Our results suggest that the regulatory mechanisms underlying the pupal transition in holometabolous insects are evolutionarily conserved in hemimetabolous G. bimaculatus, with the penultimate and final nymphal periods being equivalent to the pupal stage. This article is part of the theme issue 'The evolution of complete metamorphosis'.
Yoshiyasu Ishimaru, Tetsuya Bando, Hideyo Ohuchi, Sumihare Noji and Taro Mito : Bone morphogenetic protein signaling in distal patterning and intercalation during leg regeneration of the cricket, Gryllus bimaculatus, Development Growth & Differentiation, Vol.60, No.6, 377-386, 2018.
(Summary)
The cricket, Gryllus bimaculatus, is a classic model of leg regeneration following amputation. We previously demonstrated that Gryllus decapentaplegic (Gb'dpp) is expressed during leg regeneration, although it remains unclear whether it is essential for this process. In this study, double-stranded RNA targeting the Smad mathers-against-dpp homolog, Gb'mad, was used to examine the role of bone morphogenetic protein (BMP) signaling in the leg regeneration process of Gryllus bimaculatus. RNA interference (RNAi)-mediated knockdown of Gb'mad led to a loss of tarsus regeneration at the most distal region of regenerating leg segments. Moreover, we confirmed that the phenotype obtained by knockdown of Dpp type I receptor, Thick veins (Gb'tkv), closely resembled that observed for Gb'mad RNAi crickets, thereby suggesting that the BMP signaling pathway is indispensable for the initial stages of tarsus formation. Interestingly, knockdown of Gb'mad and Gb'tkv resulted in significant elongation of regenerating tibia along the proximodistal axis compared with normal legs. Moreover, our findings indicate that during the regeneration of tibia, the BMP signaling pathway interacts with Dachsous/Fat (Gb'Ds/Gb'Ft) signaling and dachshund (Gb'dac) to re-establish positional information and regulate determination of leg size. Based on these observations, we discuss possible roles for Gb'mad in the distal patterning and intercalation processes during leg regeneration in Gryllus bimaculatus.
Sigeo Sugano, Hiroko Suzuki, Eisuke Shimokita, Hirofumi Chiba, Sumihare Noji, Yuriko Osakabe and Keishi Osakabe : Genome editing in the mushroom-forming basidiomycetes, Coprinopsis cinerea, optimized by high-throughput transformation system., Scientific Reports, Vol.7, 2017.
(Summary)
Mushroom-forming basidiomycetes produce a wide range of metabolites and have great value not only as food but also as an important global natural resource. Here, we demonstrate CRISPR/Cas9-based genome editing in the model species Coprinopsis cinerea. Using a high-throughput reporter assay with cryopreserved protoplasts, we identified a novel promoter, CcDED1 pro , with seven times stronger activity in this assay than the conventional promoter GPD2. To develop highly efficient genome editing using CRISPR/Cas9 in C. cinerea, we used the CcDED1 pro to express Cas9 and a U6-snRNA promoter from C. cinerea to express gRNA. Finally, CRISPR/Cas9-mediated GFP mutagenesis was performed in a stable GFP expression line. Individual genome-edited lines were isolated, and loss of GFP function was detected in hyphae and fruiting body primordia. This novel method of high-throughput CRISPR/Cas9-based genome editing using cryopreserved protoplasts should be a powerful tool in the study of edible mushrooms.
Akihiro Yasue, Hitomi Kono, Munenori Habuta, Tetsuya Bando, Keita Sato, Junji Inoue, Seiichi Oyadomari, Sumihare Noji, Eiji Tanaka and Hideyo Ohuchi : Relationship between somatic mosaicism of Pax6 mutation and variable developmental eye abnormalities-an analysis of CRISPR genome-edited mouse embryos., Scientific Reports, Vol.7, No.1, 2017.
(Summary)
The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) system is a rapid gene-targeting technology that does not require embryonic stem cells. To demonstrate dosage effects of the Pax6 gene on eye formation, we generated Pax6-deficient mice with the CRISPR/Cas system. Eyes of founder embryos at embryonic day (E) 16.5 were examined and categorized according to macroscopic phenotype as class 1 (small eye with distinct pigmentation), class 2 (pigmentation without eye globes), or class 3 (no pigmentation and no eyes). Histologically, class 1 eyes were abnormally small in size with lens still attached to the cornea at E16.5. Class 2 eyes had no lens and distorted convoluted retinas. Class 3 eyes had only rudimentary optic vesicle-like tissues or histological anophthalmia. Genotyping of neck tissue cells from the founder embryos revealed somatic mosaicism and allelic complexity for Pax6. Relationships between eye phenotype and genotype were developed. The present results demonstrated that development of the lens from the surface ectoderm requires a higher gene dose of Pax6 than development of the retina from the optic vesicle. We further anticipate that mice with somatic mosaicism in a targeted gene generated by CRISPR/Cas-mediated genome editing will give some insights for understanding the complexity in human congenital diseases that occur in mosaic form.
Silvia Naomi Mitsui Akagi, Akihiro Yasue, Kiyoshi Masuda, Takuya Naruto, Yoshiyuki Minegishi, Seiichi Oyadomari, Sumihare Noji, Issei Imoto and Eiji Tanaka : Novel human mutation and CRISPR/Cas genome-edited mice reveal the importance of C-terminal domain of MSX1 in tooth and palate development., Scientific Reports, Vol.6, 2016.
(Summary)
Several mutations, located mainly in the MSX1 homeodomain, have been identified in non-syndromic tooth agenesis predominantly affecting premolars and third molars. We identified a novel frameshift mutation of the highly conserved C-terminal domain of MSX1, known as Msx homology domain 6 (MH6), in a Japanese family with non-syndromic tooth agenesis. To investigate the importance of MH6 in tooth development, Msx1 was targeted in mice with CRISPR/Cas system. Although heterozygous MH6 disruption did not alter craniofacial development, homozygous mice exhibited agenesis of lower incisors with or without cleft palate at E16.5. In addition, agenesis of the upper third molars and the lower second and third molars were observed in 4-week-old mutant mice. Although the upper second molars were present, they were abnormally small. These results suggest that the C-terminal domain of MSX1 is important for tooth and palate development, and demonstrate that that CRISPR/Cas system can be used as a tool to assess causality of human disorders in vivo and to study the importance of conserved domains in genes.
Genetically modified pigs for biomedical applications have been mainly generated using the somatic cell nuclear transfer technique; however, this approach requires complex micromanipulation techniques and sometimes increases the risks of both prenatal and postnatal death by faulty epigenetic reprogramming of a donor somatic cell nucleus. As a result, the production of genetically modified pigs has not been widely applied. We provide a simple method for CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 gene editing in pigs that involves the introduction of Cas9 protein and single-guide RNA into in vitro fertilized zygotes by electroporation. The use of gene editing by electroporation of Cas9 protein (GEEP) resulted in highly efficient targeted gene disruption and was validated by the efficient production of Myostatin mutant pigs. Because GEEP does not require the complex methods associated with micromanipulation for somatic reprogramming, it has the potential for facilitating the genetic modification of pigs.
Hiroko Awata, Ryo Wakuda, Yoshiyasu Ishimaru, Yuji Matsuoka, Kanta Terao, Satomi Katata, Yukihisa Matsumoto, Yoshitaka Hamanaka, Sumihare Noji, Taro Mito and Makoto Mizunami : Roles of OA1 octopamine receptor and Dop1 dopamine receptor in mediating appetitive and aversive reinforcement revealed by RNAi studies., Scientific Reports, Vol.6, 29696, 2016.
(Summary)
Revealing reinforcing mechanisms in associative learning is important for elucidation of brain mechanisms of behavior. In mammals, dopamine neurons are thought to mediate both appetitive and aversive reinforcement signals. Studies using transgenic fruit-flies suggested that dopamine neurons mediate both appetitive and aversive reinforcements, through the Dop1 dopamine receptor, but our studies using octopamine and dopamine receptor antagonists and using Dop1 knockout crickets suggested that octopamine neurons mediate appetitive reinforcement and dopamine neurons mediate aversive reinforcement in associative learning in crickets. To fully resolve this issue, we examined the effects of silencing of expression of genes that code the OA1 octopamine receptor and Dop1 and Dop2 dopamine receptors by RNAi in crickets. OA1-silenced crickets exhibited impairment in appetitive learning with water but not in aversive learning with sodium chloride solution, while Dop1-silenced crickets exhibited impairment in aversive learning but not in appetitive learning. Dop2-silenced crickets showed normal scores in both appetitive learning and aversive learning. The results indicate that octopamine neurons mediate appetitive reinforcement via OA1 and that dopamine neurons mediate aversive reinforcement via Dop1 in crickets, providing decisive evidence that neurotransmitters and receptors that mediate appetitive reinforcement indeed differ among different species of insects.
Yoshiyasu Ishimaru, Sayuri Tomonari, Yuji Matsuoka, Takahito Watanabe, Katsuyuki Miyawaki, Tetsuya Bando, Kenji Tomioka, Hideyo Ohuchi, Sumihare Noji and Taro Mito : TGF-β signaling in insects regulates metamorphosis via juvenile hormone biosynthesis., Proceedings of the National Academy of Sciences of the United States of America, Vol.113, No.20, 5634-5639, 2016.
(Summary)
Although butterflies undergo a dramatic morphological transformation from larva to adult via a pupal stage (holometamorphosis), crickets undergo a metamorphosis from nymph to adult without formation of a pupa (hemimetamorphosis). Despite these differences, both processes are regulated by common mechanisms that involve 20-hydroxyecdysone (20E) and juvenile hormone (JH). JH regulates many aspects of insect physiology, such as development, reproduction, diapause, and metamorphosis. Consequently, strict regulation of JH levels is crucial throughout an insect's life cycle. However, it remains unclear how JH synthesis is regulated. Here, we report that in the corpora allata of the cricket, Gryllus bimaculatus, Myoglianin (Gb'Myo), a homolog of Drosophila Myoglianin/vertebrate GDF8/11, is involved in the down-regulation of JH production by suppressing the expression of a gene encoding JH acid O-methyltransferase, Gb'jhamt In contrast, JH production is up-regulated by Decapentaplegic (Gb'Dpp) and Glass-bottom boat/60A (Gb'Gbb) signaling that occurs as part of the transcriptional activation of Gb'jhamt Gb'Myo defines the nature of each developmental transition by regulating JH titer and the interactions between JH and 20E. When Gb'myo expression is suppressed, the activation of Gb'jhamt expression and secretion of 20E induce molting, thereby leading to the next instar before the last nymphal instar. Conversely, high Gb'myo expression induces metamorphosis during the last nymphal instar through the cessation of JH synthesis. Gb'myo also regulates final insect size. Because Myo/GDF8/11 and Dpp/bone morphogenetic protein (BMP)2/4-Gbb/BMP5-8 are conserved in both invertebrates and vertebrates, the present findings provide common regulatory mechanisms for endocrine control of animal development.
Hiroko Awata, Takahito Watanabe, Yoshitaka Hamanaka, Taro Mito, Sumihare Noji and Makoto Mizunami : Knockout crickets for the study of learning and memory: Dopamine receptor Dop1 mediates aversive but not appetitive reinforcement in crickets, Scientific Reports, Vol.5, 15885, 2015.
(Summary)
Elucidation of reinforcement mechanisms in associative learning is an important subject in neuroscience. In mammals, dopamine neurons are thought to play critical roles in mediating both appetitive and aversive reinforcement. Our pharmacological studies suggested that octopamine and dopamine neurons mediate reward and punishment, respectively, in crickets, but recent studies in fruit-flies concluded that dopamine neurons mediates both reward and punishment, via the type 1 dopamine receptor Dop1. To resolve the discrepancy between studies in different insect species, we produced Dop1 knockout crickets using the CRISPR/Cas9 system and found that they are defective in aversive learning with sodium chloride punishment but not appetitive learning with water or sucrose reward. The results suggest that dopamine and octopamine neurons mediate aversive and appetitive reinforcement, respectively, in crickets. We suggest unexpected diversity in neurotransmitters mediating appetitive reinforcement between crickets and fruit-flies, although the neurotransmitter mediating aversive reinforcement is conserved. This study demonstrates usefulness of the CRISPR/Cas9 system for producing knockout animals for the study of learning and memory.
Yoshimasa Hamada, Tetsuya Bando, Taro Nakamura, Yoshiyasu Ishimaru, Taro Mito, Sumihare Noji, Kenji Tomioka and Hideyo Ohuchi : Regenerated leg segment patterns are regulated epigenetically by histone H3K27 methylation in the cricket Gryllus bimaculatus, Development, Vol.142, No.17, 2916-2927, 2015.
(Summary)
Hemimetabolous insects such as the cricket Gryllus bimaculatus regenerate lost tissue parts using blastemal cells, a population of dedifferentiated proliferating cells. The expression of several factors that control epigenetic modification is upregulated in the blastema compared with differentiated tissue, suggesting that epigenetic changes in gene expression might control the differentiation status of blastema cells during regeneration. To clarify the molecular basis of epigenetic regulation during regeneration, we focused on the function of the Gryllus Enhancer of zeste [Gb'E(z)] and Ubiquitously transcribed tetratricopeptide repeat gene on the X chromosome (Gb'Utx) homologues, which regulate methylation and demethylation of histone H3 lysine 27 (H3K27), respectively. Methylated histone H3K27 in the regenerating leg was diminished by Gb'E(z)(RNAi) and was increased by Gb'Utx(RNAi). Regenerated Gb'E(z)(RNAi) cricket legs exhibited extra leg segment formation between the tibia and tarsus, and regenerated Gb'Utx(RNAi) cricket legs showed leg joint formation defects in the tarsus. In the Gb'E(z)(RNAi) regenerating leg, the Gb'dac expression domain expanded in the tarsus. By contrast, in the Gb'Utx(RNAi) regenerating leg, Gb'Egfr expression in the middle of the tarsus was diminished. These results suggest that regulation of the histone H3K27 methylation state is involved in the repatterning process during leg regeneration among cricket species via the epigenetic regulation of leg patterning gene expression.
Yuji Matsuoka, Tetsuya Bando, Takahito Watanabe, Yoshiyasu Ishimaru, Sumihare Noji, Aleksandar Popadic and Taro Mito : Short germ insects utilize both the ancestral and derived mode of Polycomb group-mediated epigenetic silencing of Hox genes., Biology Open, Vol.4, No.6, 702-709, 2015.
(Summary)
In insect species that undergo long germ segmentation, such as Drosophila, all segments are specified simultaneously at the early blastoderm stage. As embryogenesis progresses, the expression boundaries of Hox genes are established by repression of gap genes, which is subsequently replaced by Polycomb group (PcG) silencing. At present, however, it is not known whether patterning occurs this way in a more ancestral (short germ) mode of embryogenesis, where segments are added gradually during posterior elongation. In this study, two members of the PcG family, Enhancer of zeste (E(z)) and Suppressor of zeste 12 (Su(z)12), were analyzed in the short germ cricket, Gryllus bimaculatus. Results suggest that although stepwise negative regulation by gap and PcG genes is present in anterior members of the Hox cluster, it does not account for regulation of two posterior Hox genes, abdominal-A (abd-A) and Abdominal-B (Abd-B). Instead, abd-A and Abd-B are predominantly regulated by PcG genes, which is the mode present in vertebrates. These findings suggest that an intriguing transition of the PcG-mediated silencing of Hox genes may have occurred during animal evolution. The ancestral bilaterian state may have resembled the current vertebrate mode of regulation, where PcG-mediated silencing of Hox genes occurs before their expression is initiated and is responsible for the establishment of individual expression domains. Then, during insect evolution, the repression by transcription factors may have been acquired in anterior Hox genes of short germ insects, while PcG silencing was maintained in posterior Hox genes.
J Adam Mellott, Keerthana Devarajan, E Heather Shinogle, S David Moore, Zsolt Talata, S Jennifer Laurence, Laird M Forrest, Sumihare Noji, Eiji Tanaka, Hinrich Staecker and S Michael Detamore : Nonviral Reprogramming of Human Wharton's Jelly Cells Reveals Differences Between ATOH1 Homologues., Tissue Engineering. Part A, Vol.21, No.11-12, 1795-1809, 2015.
(Summary)
The transcription factor atonal homolog 1 (ATOH1) has multiple homologues that are functionally conserved across species and is responsible for the generation of sensory hair cells. To evaluate potential functional differences between homologues, human and mouse ATOH1 (HATH1 and MATH-1, respectively) were nonvirally delivered to human Wharton's jelly cells (hWJCs) for the first time. Delivery of HATH1 to hWJCs demonstrated superior expression of inner ear hair cell markers and characteristics than delivery of MATH-1. Inhibition of HES1 and HES5 signaling further increased the atonal effect. Transfection of hWJCs with HATH1 DNA, HES1 siRNA, and HES5 siRNA displayed positive identification of key hair cell and support cell markers found in the cochlea, as well as a variety of cell shapes, sizes, and features not native to hair cells, suggesting the need for further examination of other cell types induced by HATH1 expression. In the first side-by-side evaluation of HATH1 and MATH-1 in human cells, substantial differences were observed, suggesting that the two atonal homologues may not be interchangeable in human cells, and artificial expression of HATH1 in hWJCs requires further study. In the future, this line of research may lead to engineered systems that would allow for evaluation of drug ototoxicity or potentially even direct therapeutic use.
Y Kadomura-Ishikawa, Katsuyuki Miyawaki, Akira Takahashi, Toshiya Masuda and Sumihare Noji : Light and abscisic acid independently regulated FaMYB10 in Fragaria x ananassa fruit, Planta, Vol.241, No.4, 953-965, 2015.
(Summary)
Light and ABA independently regulated anthocyanin biosynthesis via activation of FaMYB10 expression. FaMYB10 accelerated anthocyanin synthesis of pelargonidin 3-glucoside and cyanidin 3-glucoside during strawberry fruit ripening. Light is an integral factor in fruit ripening. Ripening in non-climacteric fruit is also effected by the plant hormone abscisic acid (ABA). However, how light and/or ABA regulate fruit ripening processes, such as strawberry color development remains elusive. Results of the present study showed light and ABA regulated strawberry fruit coloration via activation of FaMYB10 expression, an R2R3 MYB transcription factor. Light exposure increased FaMYB10 transcript levels, flavonoid pathway genes, and anthocyanin content. Exogenous ABA promoted FaMYB10 expression, and anthocyanin content, accompanied by increased ABA-responsive transcript levels and flavonoid pathway genes. ABA biosynthesis inhibitor treatment, and RNAi-mediated down-regulation of the ABA biosynthetic gene (9-cis epoxycarotenoid dioxygenase: FaNCED1), and ABA receptor (magnesium chelatase H subunit: FaCHLH/ABAR) showed inverse ABA effects. Furthermore, additive effects were observed in anthocyanin accumulation under combined light and ABA, indicating independent light and ABA signaling pathways. FaMYB10 down-regulation by Agrobacterium-mediated RNA interference (RNAi) in strawberry fruits showed decreased pelargonidin 3-glucoside and cyanidin 3-glucoside levels, accompanied by consistent flavonoid pathway gene expression levels. FaMYB10 over-expression showed opposite FaMYB10 RNAi phenotypes, particularly cyanidin 3-glucoside synthesis by FaMYB10, which was correlated with FaF3'H transcript levels. These data provided evidence that light and ABA promoted FaMYB10 expression, resulting in anthocyanin accumulation via acceleration of flavonoid pathway gene expression. Finally, our results suggested FaMYB10 serves a role as a signal transduction mediator from light and ABA perception to anthocyanin synthesis in strawberry fruit.
Yoshiyasu Ishimaru, Taro Nakamura, Tetsuya Bando, Yuji Matsuoka, Hideyo Ohuchi, Sumihare Noji and Taro Mito : Involvement of dachshund and Distal-less in distal pattern formation of the cricket leg during regeneration., Scientific Reports, Vol.5, 8387, 2015.
(Summary)
Cricket nymphs have the remarkable ability to regenerate a functional leg following amputation, indicating that the regenerating blastemal cells contain information for leg morphology. However, the molecular mechanisms that underlie regeneration of leg patterns remain poorly understood. Here, we analyzed phenotypes of the tibia and tarsus (three tarsomeres) obtained by knockdown with regeneration-dependent RNA interference (rdRNAi) against Gryllus dachshund (Gb'dac) and Distal-less (Gb'Dll). We found that depletion of Gb'Dll mRNA results in loss of the tarsal segments, while rdRNAi against Gb'dac shortens the tibia at the two most distal tarsomeres. These results indicate that Gb'Dll expression is indispensable for formation of the tarsus, while Gb'dac expression is necessary for elongation of the tibia and formation of the most proximal tarsomere. These findings demonstrate that mutual transcriptional regulation between the two is indispensable for formation of the tarsomeres, whereas Gb'dac is involved in determination of tibial size through interaction with Gb'ds/Gb'ft.
Akihiro Yasue, Silvia Naomi Mitsui Akagi, Takahito Watanabe, Tetsushi Sakuma, Seiichi Oyadomari, Takashi Yamamoto, Sumihare Noji, Taro Mito and Eiji Tanaka : Highly efficient targeted mutagenesis in one-cell mouse embryos mediated by the TALEN and CRISPR/Cas systems., Scientific Reports, Vol.4, 5705, 2014.
(Summary)
Since the establishment of embryonic stem (ES) cell lines, the combined use of gene targeting with homologous recombination has aided in elucidating the functions of various genes. However, the ES cell technique is inefficient and time-consuming. Recently, two new gene-targeting technologies have been developed: the transcription activator-like effector nuclease (TALEN) system, and the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) system. In addition to aiding researchers in solving conventional problems, these technologies can be used to induce site-specific mutations in various species for which ES cells have not been established. Here, by targeting the Fgf10 gene through RNA microinjection in one-cell mouse embryos with the TALEN and CRISPR/Cas systems, we produced the known limb-defect phenotypes of Fgf10-deficient embryos at the F0 generation. Compared to the TALEN system, the CRISPR/Cas system induced the limb-defect phenotypes with a strikingly higher efficiency. Our results demonstrate that although both gene-targeting technologies are useful, the CRISPR/Cas system more effectively elicits single-step biallelic mutations in mice.
Qing-Ri Jin, Yukiko Bandou, Katsuyuki Miyawaki, Yosuke Shikama, Chisato Kosugi, Nanako Aki, Makoto Funaki and Sumihare Noji : Correlation of fibroblast growth factor 21 serum levels with metabolic parameters in Japanese subjects, The Journal of Medical Investigation : JMI, Vol.61, No.1.2, 28-34, 2014.
(Summary)
Since serum level of fibroblast growth factor 21 (FGF21) has been implicated as a potential biomarker for the early detection of the metabolic syndrome and type 2 diabetes, we examined how FGF21 serum levels are correlated with metabolic parameters in Japanese subjects. FGF21 levels were analyzed by enzyme-linked immunosorbent assays. Spearman's correlation and multiple stepwise regression analyses were used to examine the relationship between serum FGF21 and other factors. A Mann-Whitney U test was performed between the normal and high groups for triglycerides and systolic blood pressure (BP) respectively. By univariate correlation analysis, serum FGF21 levels were significantly associated with triglyceride levels, systolic BP, diastolic BP, pulse pressure, body mass index (BMI), age, fasting plasma glucose (FPG) levels, and total cholesterol levels. Multiple regression analysis (adjusted for age, gender, and BMI) showed that serum FGF21 levels were independently and significantly associated with triglyceride levels and systolic BP. Serum FGF21 levels were significantly higher in subjects with high triglyceride levels and high systolic BP compared with those who had normal triglyceride levels and normal systolic BP respectively. This study found that FGF21 levels might be a biomarker for some metabolic disorders associated with metabolic syndrome.
(Keyword)
Adult / Aged / Asian Continental Ancestry Group / Biological Markers / Blood Glucose / blood pressure / Body Mass Index / Cross-Sectional Studies / Female / Fibroblast Growth Factors / Humans / Male / Metabolic Syndrome X / Middle Aged / Regression Analysis / Triglycerides
Hiroshi Yoshida, Tetsuya Bando, Taro Mito, Hideyo Ohuchi and Sumihare Noji : An extended steepness model for leg-size determination based on Dachsous/Fat trans-dimer system., Scientific Reports, Vol.4, 4335, 2014.
(Summary)
What determines organ size has been a long-standing biological question. Lawrence et al. (2008) proposed the steepness hypothesis suggesting that the protocadherin Dachsous/Fat (Ds/Ft) system may provide some measure of dimension to the cells in relation to the gradient. In this paper we extended the model as a means of interpreting experimental results in cricket leg regeneration. We assumed that (1) Ds/Ft trans-heterodimers or trans-homodimers are redistributed during cell division, and (2) growth would cease when a differential of the dimer across each cell decreases to a certain threshold. We applied our model to simulate the results obtained by leg regeneration experiments in a cricket model. The results were qualitatively consistent with the experimental data obtained for cricket legs by RNA interference methodology. Using our extended steepness model, we provided a molecular-based explanation for leg size determination even in intercalary regeneration and for organ size determination.
Takahito Watanabe, Sumihare Noji and Taro Mito : Gene knockout by targeted mutagenesis in a hemimetabolous insect, the two-spotted cricket Gryllus bimaculatus, using TALENs., Methods, Vol.69, No.1, 17-21, 2014.
(Summary)
Hemimetabolous, or incompletely metamorphosing, insects are phylogenetically basal. These insects include many deleterious species. The cricket, Gryllus bimaculatus, is an emerging model for hemimetabolous insects, based on the success of RNA interference (RNAi)-based gene-functional analyses and transgenic technology. Taking advantage of genome-editing technologies in this species would greatly promote functional genomics studies. Genome editing using transcription activator-like effector nucleases (TALENs) has proven to be an effective method for site-specific genome manipulation in various species. TALENs are artificial nucleases that are capable of inducing DNA double-strand breaks into specified target sequences. Here, we describe a protocol for TALEN-based gene knockout in G. bimaculatus, including a mutant selection scheme via mutation detection assays, for generating homozygous knockout organisms.
Y Kadomura-Ishikawa, Katsuyuki Miyawaki, Sumihare Noji and Akira Takahashi : Phototropin 2 is involved in blue light-induced anthocyanin accumulation in Fragaria x ananassa fruits, Journal of Plant Research, Vol.126, No.6, 847-857, 2013.
(Summary)
Anthocyanins are widespread, essential secondary metabolites in higher plants during color development in certain flowers and fruits. In strawberries, anthocyanins are also key contributors to fruit antioxidant capacity and nutritional value. However, the effects of different light qualities on anthocyanin accumulation in strawberry (Fragaria x ananassa, cv. Sachinoka) fruits remain elusive. In the present study, we showed the most efficient increase in anthocyanin content occurred by blue light irradiation. Light sensing at the molecular level was investigated by isolation of two phototropin (FaPHOT1 and FaPHOT2), two cryptochrome (FaCRY1 and FaCRY2), and two phytochrome (FaPHYA and FaPHYB) homologs. Expression analysis revealed only FaPHOT2 transcripts markedly increased depending on fruit developmental stage, and a corresponding increase in anthocyanin content was detected. FaPHOT2 knockdown resulted in decreased anthocyanin content; however, overexpression increased anthocyanin content. These findings suggested blue light induced anthocyanin accumulation, and FaPHOT2 may play a role in sensing blue light, and mediating anthocyanin biosynthesis in strawberry fruits. This is the first report to find a relationship between visible light sensing, and color development in strawberry fruits.
Emi Kawakami, Nobuhiko Kawai, Nao Kinouchi, Hiroyo Mori, Yutaka Ohsawa, Naozumi Ishimaru, Yoshihide Sunada, Sumihare Noji and Eiji Tanaka : Local applications of myostatin-siRNA with atelocollagen increase skeletal muscle mass and recovery of muscle function., PLoS ONE, Vol.8, No.5, 2013.
(Summary)
These data suggest local administration of Mst-siRNA/ATCOL complex could lead to skeletal muscle hypertrophy and recovery of motor disability in mCAV-3Tg mice. Therefore, ATCOL-mediated application of siRNA is a potential tool for therapeutic use in muscular atrophy diseases.
Tetsuya Bando, Yoshiyasu Ishimaru, Takuro Kida, Yoshimasa Hamada, Yuji Matsuoka, Taro Nakamura, Hideyo Ohuchi, Sumihare Noji and Taro Mito : Analysis of RNA-Seq data reveals involvement of JAK/STAT signalling during leg regeneration in the cricket Gryllus bimaculatus, Development, Vol.140, No.5, 959-964, 2013.
(Summary)
In the cricket Gryllus bimaculatus, missing distal parts of the amputated leg are regenerated from the blastema, a population of dedifferentiated proliferating cells that forms at the distal tip of the leg stump. To identify molecules involved in blastema formation, comparative transcriptome analysis was performed between regenerating and normal unamputated legs. Components of JAK/STAT signalling were upregulated more than twofold in regenerating legs. To verify their involvement, Gryllus homologues of the interleukin receptor Domeless (Gb'dome), the Janus kinase Hopscotch (Gb'hop) and the transcription factor STAT (Gb'Stat) were cloned, and RNAi was performed against these genes. Gb'dome(RNAi), Gb'hop(RNAi) and Gb'Stat(RNAi) crickets showed defects in leg regeneration. Blastema expression of Gb'cyclinE was decreased in the Gb'Stat(RNAi) cricket compared with that in the control. Hyperproliferation of blastema cells caused by Gb'fat(RNAi) or Gb'warts(RNAi) was suppressed by RNAi against Gb'Stat. The results suggest that JAK/STAT signalling regulates blastema cell proliferation during leg regeneration.
Inoue Junji, Y Ueda Yuuki, Bando Tetsuya, Taro Mito, Sumihare Noji and Hideyo Ohuchi : The expression of LIM-homeobox genes, Lhx1 and Lhx5, in the forebrain is essential for neural retina differentiation., Development Growth & Differentiation, Vol.55, No.7, 668-675, 2013.
Takahito Watanabe, Hiroshi Ochiai, Tetsushi Sakuma, W Hadley Horch, Naoya Hamaguchi, Taro Nakamura, Tetsuya Bando, Hideyo Ohuchi, Takashi Yamamoto, Sumihare Noji and Taro Mito : Non-transgenic genome modifications in a hemimetabolous insect using zinc-finger and TAL effector nucleases., Nature Communications, Vol.3, 1017-1025, 2012.
(Summary)
Hemimetabolous, or incompletely metamorphosing, insects are phylogenetically relatively basal and comprise many pests. However, the absence of a sophisticated genetic model system, or targeted gene-manipulation system, has limited research on hemimetabolous species. Here we use zinc-finger nuclease and transcription activator-like effector nuclease technologies to produce genetic knockouts in the hemimetabolous insect Gryllus bimaculatus. Following the microinjection of mRNAs encoding zinc-finger nucleases or transcription activator-like effector nucleases into cricket embryos, targeting of a transgene or endogenous gene results in sequence-specific mutations. Up to 48% of founder animals transmit disrupted gene alleles after zinc-finger nucleases microinjection compared with 17% after microinjection of transcription activator-like effector nucleases. Heterozygous offspring is selected using mutation detection assays that use a Surveyor (Cel-I) nuclease, and subsequent sibling crosses create homozygous knockout crickets. This approach is independent from a mutant phenotype or the genetic tractability of the organism of interest and can potentially be applied to manage insect pests using a non-transgenic strategy.
Katsuyuki Miyawaki, Sachi Fukuoka, Yasuko Kadomura, Hirokazu Hamaoka, Taro Mito, Hideyo Ohuchi, Wilfried Schwab and Sumihare Noji : Establishment of a novel system to elucidate the mechanisms underlying light-induced ripening of strawberry fruit with an Agrobacterium-mediated RNAi technique, Plant Biotechnology, Vol.29, 271-278, 2012.
(Summary)
Traditional methods used to study strawberry ripening-related gene function are time-consuming, and require at least 15 months from initiating the transformation experiment until the first ripe fruits are available for analysis. To accelerate data acquisition during gene function studies, we explored a transient assay method that employs an Agrobacterium-mediated RNAi (AmRNAi) technique in post-harvest strawberry fruit, Fragaria×ananassa (Fa) cv. Sachinoka, a Japanese cultivar. Our results showed that artificial white light induced strong expression of Fa chalcone synthase (Fa CHS), Fa chalcone isomerase (Fa CHI), and Fa flavonoid 3 -hydroxylase orthologues (Fa F3 H) in post-harvest fruit. Fa CHS and Fa F3 H function was subsequently examined by performing AmRNAi with post-harvest fruit. Although reduction of light-induced Fa F3 H expression by AmRNAi resulted in no significant change in anthocyanin content, reduction of Fa CHS significantly decreased anthocyanin levels, and up-regulated Fa F3 H levels. Our results are consistent with previous data indicating that while CHS is required for anthocyanin accumulation during late stage strawberry fruit maturation, Fa F3 H is not required. The novel system described here enabled gene function data to be available within 10 days of initiating the incubation period following infiltration. Therefore, we conclude our system is a valuable tool to elucidate the molecular mechanisms underlying light-induced ripening of strawberry fruit.
Akira Takagi, Kazuki Kurita, Taiki Terasawa, Taro Nakamura, Tetsuya Bando, Yoshiyuki Moriyama, Taro Mito, Sumihare Noji and Hideyo Ohuchi : Functional analysis of the role of eyes absent and sine oculis in the developing eye of the cricket Gryllus bimaculatus., Development Growth & Differentiation, Vol.54, No.2, 227-240, 2012.
(Summary)
In the cricket Gryllus bimaculatus, a hemimetabolous insect, the compound eyes begin to form in the embryo and increase 5-6 fold in size during the postembryonic development of the nymphal stage. Retinal stem cells in the anteroventral proliferation zone (AVPZ) of the nymphal eye proliferate to increase retinal progenitors, which then differentiate to form new ommatidia in the anterior region of the eye. However, mechanisms underlying this type of eye formation have not been well elucidated yet. Here, we found that the homologues of the retinal determination transcription factor genes of eyes absent (eya) and sine oculis (so) are expressed during the cricket embryonic eye formation. eya is also expressed intensely in the AVPZ of the nymphal eye. To explore their functions, we performed knockdown by RNA interference (RNAi). Knockdown of Gb'eya resulted in loss of the embryonic eye. In the nymphal eye, RNAi against Gb'eya or Gb'so impaired retinal morphology by apparently transforming cornea structures into head cuticle. These results imply that Gb'eya and Gb'so are essential for the differentiation of the retinal progenitor cells and maintaining retinal structures during eye development.
Taro Mito, Yohei Shinmyo, Kazuki Kurita, Taro Nakamura, Hideyo Ohuchi and Sumihare Noji : Ancestral functions of Delta/Notch signaling in the formation of body and leg segments in the cricket Gryllus bimaculatus., Development, Vol.138, No.17, 3823-3833, 2011.
(Summary)
Delta/Notch signaling controls a wide spectrum of developmental processes, including body and leg segmentation in arthropods. The various functions of Delta/Notch signaling vary among species. For instance, in Cupiennius spiders, Delta/Notch signaling is essential for body and leg segmentation, whereas in Drosophila fruit flies it is involved in leg segmentation but not body segmentation. Therefore, to gain further insight into the functional evolution of Delta/Notch signaling in arthropod body and leg segmentation, we analyzed the function of the Delta (Gb'Delta) and Notch (Gb'Notch) genes in the hemimetabolous, intermediate-germ cricket Gryllus bimaculatus. We found that Gb'Delta and Gb'Notch were expressed in developing legs, and that RNAi silencing of Gb'Notch resulted in a marked reduction in leg length with a loss of joints. Our results suggest that the role of Notch signaling in leg segmentation is conserved in hemimetabolous insects. Furthermore, we found that Gb'Delta was expressed transiently in the posterior growth zone of the germband and in segmental stripes earlier than the appearance of wingless segmental stripes, whereas Gb'Notch was uniformly expressed in early germbands. RNAi knockdown of Gb'Delta or Gb'Notch expression resulted in malformation in body segments and a loss of posterior segments, the latter probably due to a defect in posterior growth. Therefore, in the cricket, Delta/Notch signaling might be required for proper morphogenesis of body segments and posterior elongation, but not for specification of segment boundaries.
(Keyword)
Animals / Body Patterning / Cell Proliferation / Embryo, Nonmammalian / Embryonic Development / Extremities / Gryllidae / Immunohistochemistry / In Situ Hybridization / Intracellular Signaling Peptides and Proteins / Membrane Proteins / RNA Interference / Receptors, Notch / Signal Transduction
Noha Dabour, Tetsuya Bando, Taro Nakamura, Katsuyuki Miyawaki, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Cricket body size is altered by systemic RNAi against insulin signaling components and epidermal growth factor receptor., Development Growth & Differentiation, Vol.53, No.7, 857-869, 2011.
(Summary)
A long-standing problem of developmental biology is how body size is determined. In Drosophila melanogaster, the insulin/insulin-like growth factor (I/IGF) and target of rapamycin (TOR) signaling pathways play important roles in this process. However, the detailed mechanisms by which insect body growth is regulated are not known. Therefore, we have attempted to utilize systemic nymphal RNA interference (nyRNAi) to knockdown expression of insulin signaling components including Insulin receptor (InR), Insulin receptor substrate (chico), Phosphatase and tensin homologue (Pten), Target of rapamycin (Tor), RPS6-p70-protein kinase (S6k), Forkhead box O (FoxO) and Epidermal growth factor receptor (Egfr) and observed the effects on body size in the Gryllus bimaculatus cricket. We found that crickets treated with double-stranded RNA (dsRNA) against Gryllus InR, chico, Tor, S6k and Egfr displayed smaller body sizes, while Gryllus FoxO nyRNAi-ed crickets exhibited larger than normal body sizes. Furthermore, RNAi against Gryllus chico and Tor displayed slow growth and RNAi against Gryllus chico displayed longer lifespan than control crickets. Since no significant difference in ability of food uptake was observed between the Gryllus chico(nyRNAi) nymphs and controls, we conclude that the adult cricket body size can be altered by knockdown of expressions of Gryllus InR, chico, Tor, S6k, FoxO and Egfr by systemic RNAi. Our results suggest that the cricket is a promising model to study mechanisms underlying controls of body size and life span with RNAi methods.
Tetsuya Bando, Taro Mito, Taro Nakamura, Hideyo Ohuchi and Sumihare Noji : Regulation of leg size and shape:involvement of the Dachsous/Fat signaling pathway, Developmental Dynamics, Vol.240, No.5, 1028-1041, 2011.
(Summary)
How limb size and shape is regulated is a long-standing question in developmental and regeneration biology. Recently, the protocadherin Dachsous-Fat (Ds-Ft) signaling pathway has been found to be essential for wing development of the fly and leg regeneration of the cricket. The Ds-Ft signaling pathway is linked to the Warts-Hippo (Wts-Hpo) signaling pathway, leading to cell proliferation. Several lines of evidence have suggested that the Wts-Hpo signaling pathway is involved in the control of organ size, and that this pathway is regulated by Ds-Ft and Merlin-Expanded, which are linked to morphogens such as decapentaplegic/bone morphogenic protein, Wingless/Wnt, and epidermal growth factor. Here we review recent progress in elucidating mechanisms controlling leg size and shape in insects and vertebrates, focusing on the Ds-Ft signaling pathway. We also introduce a working model, Ds-Ft steepness model, to explain how steepness of the Ds-Ft gradient controls leg size along the proximodistal axis.
Tetsuya Bando, Yoshimasa Hamada, Kazuki Kurita, Taro Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Lowfat, a mammalian Lix1 homologue, regulates leg size and growth under the Dachsous/Fat signaling pathway during tissue regeneration., Developmental Dynamics, Vol.240, No.6, 1440-1453, 2011.
(Summary)
In the cricket Gryllus bimaculatus, missing distal parts of amputated legs are regenerated from blastemas based on positional information. The Dachsous/Fat (Ds/Ft) signaling pathway regulates blastema cell proliferation and positional information along the longitudinal axis during leg regeneration. Herein, we show that the Gryllus homologue of Lowfat (Gb'Lft), which modulates Ds/Ft signaling in Drosophila, is involved in leg regeneration. Gb'lft is expressed in regenerating legs, and RNAi against Gb'lft (Gb'lft(RNAi)) suppressed blastema cell hyperproliferation caused by Gb'ft(RNAi) or Gb'ds(RNAi) but enhanced that caused by Gb'kibra(RNAi) or Gb'warts(RNAi). In Gb'lft(RNAi) nymphs, missing parts of amputated legs were regenerated, but the length of the regenerated legs was shortened depending on the position of the amputation. Both normal and reversed intercalary regeneration occurred in Gb'lft(RNAi) nymphs, suggesting that Gb'Lft is involved in blastema cell proliferation and longitudinal leg regeneration under the Ds/Ft signaling pathway, but it is not required for intercalary regeneration.
Small interfering RNA (siRNA)-mediated silencing of gene expression is rapidly becoming a powerful tool for molecular therapy. However, the rapid degradation of siRNAs and their limited duration of activity require efficient delivery methods. Atelocollagen (ATCOL)-mediated administration of siRNAs is a promising approach to disease treatment, including muscular atrophy. Herein, we report that ATCOL-mediated systemic administration of a myostatin-targeting siRNA into a caveolin-3-deficient mouse model of limb-girdle muscular dystrophy 1C (LGMD1C) induced a marked increase in muscle mass and a significant recovery of contractile force. These results provide evidence that ATCOL-mediated systemic administration of siRNAs may be a powerful therapeutic tool for disease treatment, including muscular atrophy.
Seiichi Koike, Yoshifumi Yutoh, Kazuko Keino-Masu, Sumihare Noji, Masayuki Masu and Hideyo Ohuchi : Autotaxin is required for the cranial neural tube closure and establishment of the midbrain-hindbrain boundary during mouse development., Developmental Dynamics, Vol.240, No.2, 413-421, 2011.
(Summary)
Autotaxin (ATX) is a lysophospholipid-generating exoenzyme expressed in embryonic and adult neural tissues. We previously showed that ATX is expressed in the neural organizing centers, anterior head process, and midbrain-hindbrain boundary (MHB). To elucidate the role of ATX during neural development, here we examined the neural phenotypes of ATX-deficient mice. Expression analysis of neural marker genes revealed that lateral expansion of the rostral forebrain is reduced and establishment of the MHB is compromised as early as the late headfold stage in ATX mutant embryos. Moreover, ATX mutant embryos fail to complete cranial neural tube closure. These results indicate that ATX is essential for cranial neurulation and MHB establishment.
Taro Mito, T Nakamura, T Band, Hideyo Ohuchi and Sumihare Noji : The advent of RNA interference in Entomology, Entomological Science, Vol.14, No.1, 1-8, 2011.
Taro Adachi, Emi Kawakami, Naozumi Ishimaru, Takahiro Ochiya, Yoshio Hayashi, Hideyo Ohuchi, Masao Tanihara, Eiji Tanaka and Sumihare Noji : Delivery of small interfering RNA with a synthetic collagen poly(Pro-Hyp-Gly) for gene silencing in vitro and in vivo., Development Growth & Differentiation, Vol.52, No.8, 693-699, 2010.
(Summary)
Silencing gene expression by small interfering RNAs (siRNAs) has become a powerful tool for the genetic analysis of many animals. However, the rapid degradation of siRNA and the limited duration of its action in vivo have called for an efficient delivery technology. Here, we describe that siRNA complexed with a synthetic collagen poly(Pro-Hyp-Gly) (SYCOL) is resistant to nucleases and is efficiently transferred into cells in vitro and in vivo, thereby allowing long-term gene silencing in vivo. We found that the SYCOL-mediated local application of siRNA targeting myostatin, coding a negative regulator of skeletal muscle growth, in mouse skeletal muscles, caused a marked increase in the muscle mass within a few weeks after application. Furthermore, in vivo administration of an anti-luciferase siRNA/SYCOL complex partially reduced luciferase expression in xenografted tumors in vivo. These results indicate a SYCOL-based non-viral delivery method could be a reliable simple approach to knockdown gene expression by RNAi in vivo as well as in vitro.
(Keyword)
Animals / Base Sequence / Cell Line, Tumor / Collagen / DNA Primers / Gene Silencing / Mice / RNA, Small Interfering / Reverse Transcriptase Polymerase Chain Reaction
Hideyo Ohuchi, Hitomi Fukui, Akane Matsuyo, Sayuri Tomonari, Masayuki Tanaka, Hiroyuki Arai, Sumihare Noji and Junken Aoki : Autotaxin controls caudal diencephalon-mesencephalon development in the chick., Developmental Dynamics, Vol.239, No.10, 2647-2658, 2010.
(Summary)
The diencephalon is the embryonic anlagen of the higher integration centers of the brain. Recent studies have elucidated how the cells in the rostral diencephalon acquire their regional identities. However, the understanding of the mechanisms under which the caudal diencephalon is formed is still limited. Here we focus on the role of Autotaxin (ATX), a lysophospholipid-generating exoenzyme, whose mRNA is detected in the caudal diencephalon. RNA interference against ATX altered the expression pattern of Pax6-regualted genes, Tcf4, Lim1, and En1, implying that ATX is required for the maintenance of the regional identity of the caudal diencephalon and the diencephalon-mesencephalon boundary (DMB). Furthermore, ATX-RNAi inhibited neuroepithelial cell proliferation on both sides of the DMB. We propose a dual role of ATX in chick brain development, in which ATX not only contributes to the formation of caudal diencephalon as a short-range signal, but also regulates the growth of mesencephalon as a long-range signal.
Taro Nakamura, Masato Yoshizaki, Syoutaro Ogawa, H Okamoto, Yohei Shinmyo, Tetsuya Bando, Hideyo Ohuchi, Sumihare Noji and Taro Mito : Imaging of transgenic cricket embryos reveals cell movements consistent with a syncytial patterning mechanism, Current Biology, Vol.20, No.18, 1641-1647, 2010.
(Summary)
The mode of insect embryogenesis varies among species, reflecting adaptations to different life history strategies [1, 2]. In holometabolous insects, which include the model systems, such as the fruit fly and the red flour beetle, a large proportion of the blastoderm produces an embryo, whereas hemimetabolous embryos generally arise from a small region of the blastoderm [3]. Despite their importance in evolutionary studies, information of early developmental dynamics of hemimetabolous insects remains limited. Here, to clarify how maternal and gap gene products act in patterning the embryo of basal hemimetabolous insects, we analyzed the dynamic segmentation process in transgenic embryos of an intermediate-germ insect species, the cricket, Gryllus bimaculatus. Our data based on live imaging of fluorescently labeled embryonic cells and nuclei suggest that the positional specification of the cellular blastoderm may be established in the syncytium, where maternally derived gradients could act fundamentally in a way that is similar to that of Drosophila, namely throughout the egg. Then, the blastoderm cells move dynamically, retaining their positional information to form the posteriorly localized germ anlage. Furthermore, we find that the anterior head region of the cricket embryo is specified by orthodenticle in a cellular environment earlier than the gnathal and thoracic regions. Our findings imply that the syncytial mode of the early segmentation in long-germ insects evolved from a dynamic syncytial-to-cellular mode found in the present study, accompanied by a heterochronic shift of gap gene action.
Sumihare Noji, 足立 太郎, Emi Kawakami and Eiji Tanaka : RNA干渉法による治療を実現するための研究, The Journal of Chu-Shikoku Orthodontic Society, Vol.22, No.1, 3-8, 2010.
42.
Taro Mito, Taro Nakamura and Sumihare Noji : Evolution of insect development: to the hemimetabolous paradigm., Current Opinion in Genetics & Development, Vol.20, No.4, 355-361, 2010.
(Summary)
Mechanisms of insect development have been extensively studied in Drosophila melanogaster, a holometabolous insect. However, recent studies on other insects have gradually revealed that there exist new developmental paradigms. In this review, we focus on the new hemimetabolous paradigm. We highlight how hemimetabolous short-germ or intermediate-germ embryos establish the anterior/posterior (A/P) pattern and the importance of dynamic cell movement during germband formation. In hemimetabolous insects, orthodenticle, encoding a homeodomain-containing transcription factor, and wingless/Wnt signaling could play crucial roles in the A/P pattern formation. We also discuss recent evidence suggesting that insect developmental modes may have evolved by heterochronic shifts, while retaining certain universal metazoan features.
K Sun, N Suzuki, Z Li, R Araki, K Ueno, S Juodkazis, M Abe, Sumihare Noji and H Misawa : High-fidelity fractionation of ssDNA fragments differing in size by one-base on a spiral-channel electrophoretic chip, Electrophoresis, Vol.30, No.24, 4277-4284, 2009.
(Summary)
For the fractionation of fragments of interest from selective PCR products generated by high coverage gene expression profiling (HiCEP) analysis, high-resolution with the ability to discriminate and fractionate fragments differing by one base (base pair) in size is highly required. We report here on a new 4-inch diameter spiral-channel chip device for automatic high-fidelity fractionation. Overlapping DNA fragments of 180, 181 and 182 bases, with only one-base difference in size, were successfully fractionated. The collected fragments were PCR amplified, and then evaluated by size checking analysis, DNA sequencing, and homolog search. The high-resolution fractionation has been achieved because of the combined contributions of (i) the high-resolution separation using a 30 cm long spiral channel, (ii) a blocking technique to avoid contamination from unselected fragments during CE, and (iii) precise micro-scale target extraction. Contaminations due to unselected fractions have been greatly decreased to a negligible level by optimization of the extraction position and extraction time corresponding to the targeted segment only. This technique can be adapted to a wide range of applications, such as protein or cell collections where requirements for the high purity are more important than the amount of recovered fractionated material.
A Hamada, K Miyawaki, E Honda-sumi, K Tomioka, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Loss-of-function analyses of the fragile X-related and dopamine receptor genes by RNA interference in the cricket Gryllus bimaculatus, Developmental Dynamics, Vol.238, No.8, 2025-2033, 2009.
(Summary)
In order to explore a possibility that the cricket Gryllus bimaculatus would be a useful model to unveil molecular mechanisms of human diseases, we performed loss-of-function analyses of Gryllus genes homologous to human genes that are responsible for human disorders, fragile X mental retardation 1 (fmr1) and Dopamine receptor (DopR). We cloned cDNAs of their Gryllus homologues, Gb'fmr1, Gb'DopRI, and Gb'DopRII, and analyzed their functions with use of nymphal RNA interference (RNAi). For Gb'fmr1, three major phenotypes were observed: (1) abnormal wing postures, (2) abnormal calling song, and (3) loss of the circadian locomotor rhythm, while for Gb'DopRI, defects of wing posture and morphology were found. These results indicate that the cricket has the potential to become a novel model system to explore human neuronal pathogenic mechanisms and to screen therapeutic drugs by RNAi.
T Bando, Taro Mito, Y Maeda, T Nakamura, F Ito, T Watanabe, Hideyo Ohuchi and Sumihare Noji : Regulation of leg size and shape by the Dachsous/Fat signalling pathway during regeneration, Development, Vol.136, No.13, 2235-2245, 2009.
(Summary)
An amputated cricket leg regenerates all missing parts with normal size and shape, indicating that regenerating blastemal cells are aware of both their position and the normal size of the leg. However, the molecular mechanisms regulating this process remain elusive. Here, we use a cricket model to show that the Dachsous/Fat (Ds/Ft) signalling pathway is essential for leg regeneration. We found that knockdown of ft or ds transcripts by regeneration-dependent RNA interference (rdRNAi) suppressed proliferation of the regenerating cells along the proximodistal (PD) axis concomitantly with remodelling of the pre-existing stump, making the regenerated legs shorter than normal. By contrast, knockdown of the expanded (ex) or Merlin (Mer) transcripts induced over-proliferation of the regenerating cells, making the regenerated legs longer. These results are consistent with those obtained using rdRNAi during intercalary regeneration induced by leg transplantation. We present a model to explain our results in which the steepness of the Ds/Ft gradient controls growth along the PD axis of the regenerating leg.
(Keyword)
Animals / Cadherins / Cell Adhesion Molecules / Extremities / Gryllidae / In Situ Hybridization / Insect Proteins / Molecular Sequence Data / Neurofibromin 2 / Regeneration / Signal Transduction
M Okamoto, T Bito, Sumihare Noji and Hideyo Ohuchi : Subtype-specific expression of Fgf19 during horizontal cell development of the chicken retina, Gene Expression Patterns, Vol.9, No.5, 306-313, 2009.
(Summary)
The mechanisms underlying retinal cell diversification are crucial to proper neural development. Fibroblast growth factor 19 (Fgf19) is expressed by developing horizontal cells (HCs) in the chicken retina. Although there are two major HC subtypes, axon-bearing and axon-less, the precise subtype expressing Fgf19 remains uncertain. Here we characterize Fgf19-expressing cells by co-labeling with antibodies against Lim1 (LIM homeodomain 1, or Lhx1), Islet1, and Prox1 (prospero-related homeobox 1) which are axon-bearing HC, axon-less HC, and pan-HC markers, respectively. We found that a subset of Fgf19-expressing cells was positive for Prox1 and Lim1 in the vitread neuroepithelium at embryonic day 4 (E4). By E9, the majority of Fgf19-expressing cells became positive for Prox1 and Lim1 prior to arrival at the prospective HC layer. In contrast, Fgf19-expressing cells did not overlap with the Islet1-positive population at any stage examined. These results suggest that Fgf19 is expressed by the early migratory horizontal precursors, and later by the presumptive axon-bearing HCs.
T Takahashi, A Hamada, K Miyawaki, Y Matsumoto, Taro Mito, Sumihare Noji and M Mizunami : Systemic RNA interference for the study of learning and memory in an insect, Journal of Neuroscience Methods, Vol.179, No.1, 9-15, 2009.
(Summary)
RNA interference (RNAi) is a powerful technique for the study of molecular mechanisms underlying many biological processes, including brain functions. Among methods for RNAi, systemic administration of double-stranded RNA (systemic RNAi) is the most convenient for basic research as well as medical application, but it has yielded only limited success. To our knowledge, systemic RNAi has not been achieved for the study of learning and memory in any animals. Here we demonstrate successful systemic RNAi of the NOS gene coding for nitric oxide synthase, which, as we previously suggested, plays a critical role in the formation of olfactory long-term memory (LTM), in the nymphal cricket Gryllus bimaculatus. In situ hybridization demonstrated a high level of expression of NOS in a subset of Kenyon cells of the mushroom body, which is known to participate in olfactory learning and memory, in addition to some neurons around the antenna lobe and the base of the optic lobe. Injection of NOS double-stranded RNA (dsRNA) into the haemolymph completely impaired 1-day memory retention, although 30 min retention was unaffected. This impairment was fully rescued by injection of an NO donor, NOR3, thus suggesting that the effect of NOS dsRNA is through inhibition of NOS. Inhibition of NOS had no effects on recall of LTM. The results demonstrate that silencing of NOS expression by systemic RNAi impairs LTM formation. Systemic RNAi will become a useful method for study of the molecular mechanisms of learning and memory.
Y Moriyama, T Sakamoto, A Matsumoto, Sumihare Noji and K Tomioka : Functional analysis of the circadian clock gene period by RNA interference in nymphal crickets Gryllus bimaculatus, Journal of Insect Physiology, Vol.55, No.2, 183-187, 2009.
(Summary)
The circadian clock gene period (Gryllus bimaculatus period, Gb'per) plays a core role in circadian rhythm generation in adults of the cricket Gryllus bimaculatus. We examined the role of Gb'per in nymphal crickets that show a diurnal rhythm rather than the nocturnal rhythm of the adults. As in the adult optic lobes, Gb'per mRNA levels in the head of the third instar nymphs showed daily cycling in light-dark cycles with a peak at mid night, and the rhythm persisted in constant darkness. Injection of Gb'per double-stranded RNA (dsRNA) into the abdomen of third instar nymphs knocked-down the mRNA levels to 25% of that in control animals. Most Gb'per dsRNA injected nymphs lost their circadian locomotor activity rhythm, while those injected with DsRed2 dsRNA as a negative control clearly maintained the rhythm. These results suggest that nymphs and adults share a common endogenous clock mechanism involving the clock gene Gb'per.
(Keyword)
Animals / Behavior, Animal / Circadian Rhythm / DNA Primers / Gryllidae / Intracellular Signaling Peptides and Proteins / Nymph / Period Circadian Proteins / RNA Interference / RNA, Messenger / Time Factors
A Urasaki, Taro Mito, Sumihare Noji, R Ueda and K Kawakami : Transposition of the vertebrate Tol2 transposable element in Drosophila melanogaster, Gene, Vol.425, No.1-2, 64-68, 2008.
(Summary)
The Tol2 element is a transposon found from a genome of a vertebrate, a small teleost medaka fish. Tol2 encodes a gene for a transposase which is active in vertebrate animals so far tested; for instance, in fish, frog, chicken and mammals, and transgenesis methods using Tol2 have been developed in these model vertebrates. However, it has not been known whether Tol2 can transpose in animals other than vertebrates. Here we report transposition of Tol2 in an invertebrate Drosophila melanogaster. First, we injected a transposon donor plasmid containing a Tol2 construct and mRNA encoding the Tol2 transposase into Drosophila eggs, and found that the Tol2 construct could be excised from the plasmid. Second, we crossed the injected flies, raised the offspring, and found that the Tol2 construct was integrated into the genome of germ cells and transmitted to the next generation. Finally, we constructed a Tol2 construct containing the white gene and injected the transposon donor plasmid and the transposase mRNA into fertilized eggs from the white mutant. We analyzed their offspring, and found that G1 flies with wild type red eyes could be obtained from 35% of the injected fly. We cloned and sequenced 34 integration loci from these lines and showed that these insertions were indeed created through transposition and distributed throughout the genome. Our present study demonstrates that the medaka fish Tol2 transposable element does not require vertebrate-specific host factors for its transposition, and also provides a possibility that Tol2 may be used as a new genetic tool for transgenesis and genome analysis in Drosophila.
(Keyword)
Animals / Cloning, Molecular / DNA Transposable Elements / Drosophila melanogaster / Gene Transfer Techniques / Oryzias / Transfection
Hideyo Ohuchi, A Hamada, H Matsuda, A Takagi, M Tanaka, J Aoki, H Arai and Sumihare Noji : Expression patterns of the lysophospholipid receptor genes during mouse early development, Developmental Dynamics, Vol.237, No.11, 3280-3294, 2008.
(Summary)
Lysophospholipids (LPs) such as lysophosphatidic acid (LPA) and sphingosine-1-phosphate (S1P) are known to mediate various biological responses, including cell proliferation, migration, and differentiation. To better understand the role of these lipids in mammalian early development, we applied whole-mount in situ hybridization techniques to E8.5 to E12.5 mouse embryos. We determined the expression patterns of the following LP receptor genes, which belong to the G protein-coupled receptor (GPCR) family: EDG1 to EDG8 (S1P1 to S1P5 and LPA1 to LPA3), LPA4 (GPR23/P2Y9), and LPA5 (GPR92). We found that the S1P/LPA receptor genes exhibit overlapping expression patterns in a variety of organ primordia, including the developing brain and cardiovascular system, presomitic mesoderm and somites, branchial arches, and limb buds. These results suggest that multiple receptor systems for LPA/S1P lysophospholipids may be functioning during organogenesis.
Naozumi Ishimaru, Rieko Arakaki, Satoko Yoshida, Akiko Yamada, Sumihare Noji and Yoshio Hayashi : Expression of the retinoblastoma protein RbAp48 in exocrine glands leads to Sjögren's syndrome-like autoimmune exocrinopathy., The Journal of Experimental Medicine, Vol.205, No.12, 2915-2927, 2008.
(Summary)
Although several autoimmune diseases are known to develop in postmenopausal women, the mechanisms by which estrogen deficiency influences autoimmunity remain unclear. Recently, we found that retinoblastoma-associated protein 48 (RbAp48) induces tissue-specific apoptosis in the exocrine glands depending on the level of estrogen deficiency. In this study, we report that transgenic (Tg) expression of RbAp48 resulted in the development of autoimmune exocrinopathy resembling Sjögren's syndrome. CD4(+) T cell-mediated autoimmune lesions were aggravated with age, in association with autoantibody productions. Surprisingly, we obtained evidence that salivary and lacrimal epithelial cells can produce interferon-gamma (IFN-gamma) in addition to interleukin-18, which activates IFN regulatory factor-1 and class II transactivator. Indeed, autoimmune lesions in Rag2(-/-) mice were induced by the adoptive transfer of lymph node T cells from RbAp48-Tg mice. These results indicate a novel immunocompetent role of epithelial cells that can produce IFN-gamma, resulting in loss of local tolerance before developing gender-based autoimmunity.
Kai Sun, Nobuko Suzuki, Zheyu Li, Ryoko Araki, Kosei Ueno, Saulius Juodkazis, Masumi Abe, Sumihare Noji and Hiroaki Misawa : Electrophoretic chip for fractionation of selective DNA fragment, Electrophoresis, Vol.29, No.19, 3959-3963, 2008.
(Summary)
A microchannel chip has been used to fractionate selected segments from an electrophoretic flow of separated fragments. A sample, which covers the size from 35 to 670 bp, was initially separated using an 8.8-cm-long channel at the electric field strength of 100 V/cm. The target fragment of 318 bp was selected and extracted from the separation channel. High-resolution fractionation was achieved by introducing new procedures for blocking, extraction, and segment transfer. Fractionation quality with and without blocking were compared using a 310 Genetic Analyzer (Applied Biosystems). The results show that no contamination was found in the sample, which was fractionated with blocking; however, a contamination by short segments was found in the sample, which was fractionated without blocking. Furthermore, fractionation by the chip was found to be of higher fidelity than that by the polyacrylamide slab gel, which displayed a small overlapped peak after the target peak. Compared with the traditional method, our chips enable faster and high-fidelity fractionation, thus providing a new tool for bioanalysis and other applications.
(Keyword)
Acrylamides / Chemical Fractionation / DNA / DNA, Single-Stranded / Electrophoresis, Microchip
Y Moriyama, T Sakamoto, SG Karpova, A Matsumoto, Sumihare Noji and K Tomioka : RNA interference of the clock gene period disrupts circadian rhythms in the cricket Gryllus bimaculatus, Journal of Biological Rhythms, Vol.23, No.4, 308-318, 2008.
(Summary)
Periodic expression of so-called clock genes is an essential part of the circadian clock. In Drosophila melanogaster the cyclic expression of per and tim through an autoregulatory feedback loop is believed to play a central role in circadian rhythm generation. However, it is still elusive whether this hypothesis is applicable to other insect species. Here it is shown that per gene plays a key role in the rhythm generation in the cricket Gryllus bimaculatus. Measurement of per mRNA levels in the optic lobe revealed the rhythmic expression of per in light cycles with a peak in the late day to early night, persisting in constant darkness. A single injection of per double-stranded RNA (dsRNA) into the abdomen of the final instar nymphs effectively knocked down the mRNA levels as adult to about 50% of control animals. Most of the per dsRNA-injected crickets completely lost the circadian locomotor activity rhythm in constant darkness up to 50 days after the injection, whereas those injected with DsRed2 dsRNA as a negative control clearly maintained it. The electrical activity of optic lobe efferents also became arrhythmic in the per dsRNA-injected crickets. These results not only suggest that per plays an important role in the circadian rhythm generation also in the cricket but also show that RNA interference is a powerful tool to dissect the molecular machinery of the cricket circadian clock.
T Nakamura, Taro Mito, K Miyawaki, Hideyo Ohuchi and Sumihare Noji : EGFR signaling is required for re-establishing the proximodistal axis during distal leg regeneration in the cricket Gryllus bimaculatus nymph, Developmental Biology, Vol.319, No.1, 46-55, 2008.
(Summary)
Nymphs of hemimetabolous insects, such as cockroaches and crickets, possess functional legs with a remarkable capacity for epimorphic regeneration. In this study, we have focused on the role of epidermal growth factor receptor (EGFR) signaling in regeneration of a nymphal leg in the cricket Gryllus bimaculatus. We performed loss-of-function analyses with a Gryllus Egfr homolog (Gb'Egfr) and nymphal RNA interference (RNAi). After injection of double-stranded RNA for Gb'Egfr in the body cavity of the third instar cricket nymph, amputation of the leg at the distal tibia resulted in defects of normal distal regeneration. The regenerated leg lacked the distal tarsus and pretarsus. This result indicates that EGFR signaling is required for distal leg patterning in regeneration during the nymphal stage of the cricket. Furthermore, we demonstrated that EGFR signaling acts downstream of the canonical Wnt/Wg signaling and regulates appendage proximodistal (PD) patterning genes aristaless and dachshund during regeneration. Our results suggest that EGFR signaling influences positional information along the PD axis in distal leg patterning of insects, regardless of the leg formation mode.
Taro Mito, Taro Nakamura, Isao Sarashina, CC Chang, S Ogawa, Hideyo Ohuchi and Sumihare Noji : Dynamic expression patterns of vasa during embryogenesis in the cricket Gryllus bimaculatus, Development Genes and Evolution, Vol.218, No.7, 381-387, 2008.
(Summary)
The specification of germ cells during embryogenesis is an important issue in the development of metazoans. In insects, the mode of germ cell specification appears to be highly variable among species and molecular data are not sufficient to provide an evolutionary perspective to this issue. Expression of vasa can be used as a germ line marker. Here, we report the isolation of a vasa-like gene in a hemimetabolous insect, the cricket Gryllus bimaculatus (Gb'vas), and its expression patterns during oogenesis and embryogenesis. Gb'vas is preferentially expressed in the germarium and the expression of Gb'vas is detectable throughout vitellogenesis including mature eggs subjected to oviposition, suggesting that Gb'vas is maternally contributed to the cricket eggs. The zygotic expression of Gb'vas appears to start at the mid blastoderm stage in the posterior region of the egg, expanding in a developing germ anlage. In early germbands, an intense expression of Gb'vas is restricted to the posterior end. In later embryos, Gb'vas expression extends over the whole body and then distinctly localized to the embryonic gonad at the stage immediately before hatching. These results suggest that, in the cricket, germ cells are specified early in development at the posterior end of an early germband, as proposed by Heymons (1895) based on cytological criteria.
Sayuri Tomonari, K Migita, A Takagi, Sumihare Noji and Hideyo Ohuchi : Expression patterns of the opsin 5-related genes in the developing chicken retina, Developmental Dynamics, Vol.237, No.7, 1910-1922, 2008.
(Summary)
The opsin gene family encodes G protein-coupled seven-transmembrane proteins that bind to a retinaldehyde chromophore for photoreception. It has been reported that opsin 5 is expressed in mammalian neural tissue, but its function has been elusive. As a first step to understand the function for opsin 5 in the developing eye, we searched for chicken opsin 5-related genes in the genome by a bioinformatic approach and isolated opsin 5 cDNA fragments from the embryonic retina by RT-PCR. We found that there are three opsin 5-related genes, designated cOpn5m (chicken opsin 5, mammalian type), cOpn5L1 (chicken opsin 5-like 1), and cOpn5L2 (chicken opsin 5-like 2), in the chicken genome. Quantitative PCR analysis has revealed that cOpn5m is the most abundant in the developing and early posthatching neural retina. In situ hybridization analysis has shown that cOpn5m is specifically expressed in subsets of differentiating ganglion cells and amacrine cells. These results suggest that the mammalian type opsin 5 may contribute to the development of these retinal cells in the chicken.
Y Ohsawa, T Okada, A Kuga, S Hayashi, T Murakami, K Tsuchida, Sumihare Noji and Y Sunada : Caveolin-3 regulates myostatin signaling. Mini-review, Acta Myologica : Myopathies and Cardiomyopathies, Vol.27, 19-24, 2008.
(Summary)
Caveolins, components of the uncoated invaginations of plasma membrane, regulate signal transduction and vesicular trafflicking. Loss of caveolin-3, resulting from dominant negative mutations of caveolin-3 causes autosomal dominant limb-girdle muscular dystrophy (LGMD) 1C and autosomal dominant rippling muscle disease (AD-RMD). Myostatin, a member of the muscle-specific transforming growth factor (TGF)-beta superfamily, negatively regulates skeletal muscle volume. Herein we review caveolin-3 suppressing of activation of type I myostatin receptor, thereby inhibiting subsequent intracellular signaling. In addition, a mouse model of LGMD1C has shown atrophic myopathy with enhanced myostatin signaling. Myostatin inhibition ameliorates muscular phenotype in the model mouse, accompanied by normalized myostatin signaling. Enhanced myostatin signaling by caveolin-3 mutation in human may contribute to the pathogenesis of LGMD1C. Therefore, myostatin inhibition therapy may be a promising treatment for patients with LGMD1C. More recent studies concerning regulation of TGF-beta superfamily signaling by caveolins have provided new insights into the pathogenesis of several human diseases.
Nao Kinouchi, Yutaka Ohsawa, Naozumi Ishimaru, Hideyo Ohuchi, Y Sunada, Yoshio Hayashi, Yukiho Tanimoto, Keiji Moriyama and Sumihare Noji : Atelocollagen-mediated local and systemic applications of myostatin-targeting siRNA increase skeletal muscle mass., Gene Therapy, Vol.15, No.15, 1126-1130, 2008.
(Summary)
RNA interference (RNAi) offers a novel therapeutic strategy based on the highly specific and efficient silencing of a target gene. Since it relies on small interfering RNAs (siRNAs), a major issue is the delivery of therapeutically active siRNAs into the target tissue/target cells in vivo. For safety reasons, strategies based on vector delivery may be of only limited clinical use. The more desirable approach is to directly apply active siRNAs in vivo. Here, we report the effectiveness of in vivo siRNA delivery into skeletal muscles of normal or diseased mice through nanoparticle formation of chemically unmodified siRNAs with atelocollagen (ATCOL). ATCOL-mediated local application of siRNA targeting myostatin, a negative regulator of skeletal muscle growth, in mouse skeletal muscles or intravenously, caused a marked increase in the muscle mass within a few weeks after application. These results imply that ATCOL-mediated application of siRNAs is a powerful tool for future therapeutic use for diseases including muscular atrophy.
Mayumi Okamoto, Sayuri Tomonari, Yuki Naito, Kaoru Saigo, Sumihare Noji, Kumiko Ui-Tei and Hideyo Ohuchi : Introduction of silencing-inducing transgene against Fgf19 does not affect expression of Tbx5 and beta3-tubulin in the developing chicken retina., Development Growth & Differentiation, Vol.50, No.3, 159-168, 2008.
(Summary)
Fgf19 is known to be expressed in the developing chicken eye but its functions during retinal development have remained elusive. Since Fgf19 is expressed in the dorsal portion of the optic cup, it is intriguing to know whether FGF19 is required for expression of dorso-ventral morphogenetic genes in the eye. To clarify this, expression patterns of Tbx5 and Vax were examined in the developing eye after in ovo RNA interference targeted against Fgf19. Quantitative polymerase chain reaction (PCR) analysis showed that the short-hairpin RNAs (shRNAs) targeted against Fgf19 could reduce its expression in the eye to less than 50% of a relative amount of mRNA, compared with contralateral or untreated control eyes. However, no obvious alteration in expression domains of Tbx5 or Vax was observed. Misexpression of Tbx5 or Tbx5-RNAi did not alter the Fgf19 expression either. Furthermore, although Fgf19 is expressed in the central retina before neurogenesis occurs, beta3-tubulin, a marker for early retinal differentiation was still detected in the central retina after knockdown of Fgf19. Thus, knockdown of Fgf19 supports no obvious regulations between Fgf19 and Tbx5, or exhibits no phenotypes that perturb early retinal differentiation.
Masashi Nakatani, Yuka Takehara, Hiromu Sugino, Mitsuru Matsumoto, Osamu Hashimoto, Yoshihisa Hasegawa, Tatsuya Murakami, Akiyoshi Uezumi, Shin'ichi Takeda, Sumihare Noji, Yoshihide Sunada and Kunihiro Tsuchida : Transgenic expression of a myostatin inhibitor derived from follistatin increases skeletal muscle mass and ameliorates dystrophic pathology in mdx mice., The FASEB journal, Vol.22, No.2, 477-487, 2008.
(Summary)
Myostatin is a potent negative regulator of skeletal muscle growth. Therefore, myostatin inhibition offers a novel therapeutic strategy for muscular dystrophy by restoring skeletal muscle mass and suppressing the progression of muscle degeneration. The known myostatin inhibitors include myostatin propeptide, follistatin, follistatin-related proteins, and myostatin antibodies. Although follistatin shows potent myostatin-inhibiting activities, it also acts as an efficient inhibitor of activins. Because activins are involved in multiple functions in various organs, their blockade by follistatin would affect multiple tissues other than skeletal muscles. In the present study, we report the characterization of a myostatin inhibitor derived from follistatin, which does not affect activin signaling. The dissociation constants (K(d)) of follistatin to activin and myostatin are 1.72 nM and 12.3 nM, respectively. By contrast, the dissociation constants (K(d)) of a follistatin-derived myostatin inhibitor, designated FS I-I, to activin and myostatin are 64.3 microM and 46.8 nM, respectively. Transgenic mice expressing FS I-I, under the control of a skeletal muscle-specific promoter showed increased skeletal muscle mass and strength. Hyperplasia and hypertrophy were both observed. We crossed FS I-I transgenic mice with mdx mice, a model for Duchenne muscular dystrophy. Notably, the skeletal muscles in the mdx/FS I-I mice showed enlargement and reduced cell infiltration. Muscle strength is also recovered in the mdx/FS I-I mice. These results indicate that myostatin blockade by FS I-I has a therapeutic potential for muscular dystrophy.
Taro Mito, Monica Ronco, Tomohiro Uda, Taro Nakamura, Hideyo Ohuchi and Sumihare Noji : Divergent and conserved roles of extradenticle in body segmentation and appendage formation, respectively, in the cricket Gryllus bimaculatus., Developmental Biology, Vol.313, No.1, 67-79, 2008.
(Summary)
The cricket Gryllus bimaculatus is a typical hemimetabolous intermediate germ insect, in which the processes of segmentation and appendage formation differ from those in Drosophila, a holometabolous long germ insect. In order to compare their developmental mechanisms, we have focused on Gryllus orthologs of the Drosophila developmental regulatory genes and studied their functions. Here, we report a functional analysis of the Gryllus ortholog of extradenticle (Gb'exd) using embryonic and parental RNA interference (RNAi) techniques. We found the following: (1) RNAi suppression of Gb'exd results in the deletion or fusion of body segments. Especially the head was often very severely affected. This gap-like phenotype may be related to reduced expression of the gap genes hunchback and Krüppel in early RNAi germbands. (2) In the appendages, several segments (podomeres) were fused. (3) Head appendages including the antenna were transformed to a leg-like structure consisting of at least one proximal podomere as well as several tarsomeres. The defects in appendages are reminiscent of the phenotype caused by large exd clones in Drosophila antennal discs. These findings led us to the conclusion that (1) Gb'exd is required for segment patterning in the gnathal to abdominal region, acting in a gap gene-like manner in the anterior region. (2) Gb'exd plays important roles in formation of the appendages and the determination of their identities, acting as a regulatory switch that chooses between the fates of head appendages versus the appendage ground state. Although functions of Gb'exd in appendage patterning appear fundamentally conserved between Gryllus and Drosophila, its role in body segmentation may differ from that of Drosophila exd.
Monica Ronco, Tomohiro Uda, Taro Mito, Alessandro Minelli, Sumihare Noji and Martin Klingler : Antenna and all gnathal appendages are similarly transformed by homothorax knock-down in the cricket Gryllus bimaculatus., Developmental Biology, Vol.313, No.1, 80-92, 2008.
(Summary)
Our understanding of the developmental mechanisms underlying the vast diversity of arthropod appendages largely rests on the peculiar case of the dipteran Drosophila melanogaster. In this insect, homothorax (hth) and extradenticle (exd) together play a pivotal role in appendage patterning and identity. We investigated the role of the hth homologue in the cricket Gryllus bimaculatus by parental RNA interference. This species has a more generalized morphology than Oncopeltus fasciatus, the one other insect besides Drosophila where homothorax function has been investigated. The Gryllus head appendages represent the morphologically primitive state including insect-typical mandibles, maxillae and labium, structures highly modified or missing in Oncopeltus and Drosophila. We depleted Gb'hth function through parental RNAi to investigate its requirement for proper regulation of other appendage genes (Gb'wingless, Gb'dachshund, Gb'aristaless and Gb'Distalless) and analyzed the terminal phenotype of Gryllus nymphs. Gb'hth RNAi nymphs display homeotic and segmentation defects similar to hth mutants or loss-of-function clones in Drosophila. Intriguingly, however, we find that in Gb'hth RNAi nymphs not only the antennae but also all gnathal appendages are homeotically transformed, such that all head appendages differentiate distally as legs and proximally as antennae. Hence, Gb'hth is not specifically required for antennal fate, but fulfills a similar role in the specification of all head appendages. This suggests that the role of hth in the insect antenna is not fundamentally different from its function as cofactor of segment-specific homeotic genes in more posterior segments.
T Nakamura, Taro Mito, T Bando, Hideyo Ohuchi and Sumihare Noji : Dissecting insect leg regeneration through RNA interference., Cellular and Molecular Life Sciences, Vol.65, No.1, 64-72, 2008.
(Summary)
Nymphs of hemimetabolous insects such as cockroaches and crickets exhibit a remarkable capacity for regenerating complex structures from damaged legs. Until recent years, however, approaches to elucidate the molecular mechanisms underlying the leg regeneration process have been lacking. Taking the cricket Gryllus bimaculatus as a model, we found that a phenotype related to regeneration frequently appears during leg regeneration, even though no phenotype is induced by RNA interference (RNAi) in the cricket nymph, designated as regeneration-dependent RNAi. Since then, we have investigated the functions of various genes encoding signaling factors and cellular adhesion proteins like Fat and Dachsous during leg regeneration. In this review, we summarize the classical knowledge about insect leg regeneration and introduce recent advances concerning the signaling cascades required for regenerating a leg. Our results provide clues to the mechanisms of regeneration which are relevant to vertebrate systems.
M Mitsumori, T Adachi, K Takayanagi, Taro Mito, Hideyo Ohuchi, S Kimura, M Kokubo, T Higuchi and Sumihare Noji : Film tomography as a tool for three-dimensional image construction and gene expression studies., Development Growth & Differentiation, Vol.49, No.7, 583-589, 2007.
(Summary)
In order to observe three-dimensional (3D) expression patterns of genes in whole animals, whole organs, or whole tissues, in situ hybridization (ISH) of many sections must be carried out and then used to construct a 3D image. For this purpose, we have developed an automatic microtome to prepare tissue sections with an adhesive film. We used commercially available film suitable for sectioning and ISH. We constructed a microtome and, after adherence of the film to a paraffin-embedded tissue block, cut the block with a blade to prepare sections on film. Then, the sections-on-film were automatically set in a plastic frame that was the same size as a conventional glass slide. With this automatic microtome, tissue sections can be made for ISH or immunohistochemistry in addition to conventional hematoxylin and eosin staining without specific training. We demonstrate that we can construct 3D images of gene expression patterns obtained by ISH on sections prepared with this automatic microtome. We have designated this method as 'Film Tomography (FITO)'.
(Keyword)
Animals / Gene Expression / In Situ Hybridization / Tomography
Sayuri Tomonari, Akira Takagi, Sumihare Noji and Hideyo Ohuchi : Expression pattern of the melanopsin-like (cOpn4m) and VA opsin-like genes in the developing chicken retina and neural tissues., Gene Expression Patterns, Vol.7, No.7, 746-753, 2007.
(Summary)
We examined the expression pattern of melanopsin-like (cOpn4m) and VA opsin-like (cVAL) genes during chicken development. Two types of cOpn4m transcripts, distinct in their carboxyl terminals were found, as is the case for the chicken melanopsin (cOpn4) reported previously. The expression of cOpn4m was restricted to the developing retina, specifically to a subset of developing amacrine cells from embryonic day 10. VA opsin is one of the non-canonical opsins, reported to exist in fish so far. In this study, an aberrant type of VA opsin-like (cVAL) cDNA was isolated from chicken embryonic neural tissues. The expression of cVAL was observed in the ventral region of the developing brain and neural tube; however, specific signals for cVAL could not be detected in the developing retina. These results indicate that the additional melanopsin in avian identifies a subset of developing amacrine cells in the retina and that the aberrant transcript of the VA opsin-like gene are present during neural tube development in the chicken.
(Keyword)
Amino Acid Sequence / Animals / Base Sequence / Chick Embryo / Chickens / Gene Expression Profiling / Gene Expression Regulation, Developmental / Molecular Sequence Data / Nervous System / Neural Crest / Retina / Rod Opsins / Sequence Homology, Amino Acid / Tissue Distribution
Y Takehara-Kasamatsu, K Tsuchida, M Nakatani, T Murakami, A Kurisaki, O Hashimoto, Hideyo Ohuchi, H Kurose, K Mori, S Kagami, Sumihare Noji and H Sugino : Characterization of follistatin-related gene as a negative regulatory factor for activin family members during mouse heart development, The Journal of Medical Investigation : JMI, Vol.54, No.3-4, 276-288, 2007.
(Summary)
Follistatin-related gene (FLRG) encodes a secretory glycoprotein that has characteristic cysteine-rich follistatin domains. FLRG protein binds to and neutralizes several transforming growth factor-beta (TGF-beta) superfamily members, including myostatin (MSTN), which is a potent negative regulator of skeletal muscle mass. We have previously reported that FLRG was abundantly expressed in fetal and adult mouse heart. In this study, we analyzed the expression of FLRG mRNA during mouse heart development. FLRG mRNA was continuously expressed in the embryonic heart, whereas it was very low in skeletal muscles. By contrast, MSTN mRNA was highly expressed in embryonic skeletal muscles, whereas the expression of MSTN mRNA was rather low in the heart. In situ hybridization and immunohistochemical analysis revealed that FLRG expressed in smooth muscle of the aorta and pulmonary artery, valve leaflets of mitral and tricuspid valves, and cardiac muscles in the ventricle of mouse embryonic heart. However, MSTN was expressed in very limited areas, such as valve leaflets of pulmonary and aortic valves, the top of the ventricular and atrial septa. Interestingly, the expression of MSTN was complementary to that of FLRG, especially in the valvular apparatus. Biochemical analyses with surface plasmon resonance biosensor and reporter assays demonstrated that FLRG hardly dissociates from MSTN and activin once it bound to them, and efficiently inhibits these activities. Our results suggest that FLRG could function as a negative regulator of activin family members including MSTN during heart development.
K Sun, Z Li, K Ueno, S Juodkazis, Sumihare Noji and H Misawa : Electrophoretic chip for high-fidelity fractionation of double-stranded DNA, Electrophoresis, Vol.28, No.10, 1572-1578, 2007.
(Summary)
We report the high fidelity, on-chip fractionation of selected segments from an electrophoretic flow of separated fragments. dsDNA fragments (10-330 base pairs (bp)) were initially separated using a 6.5 cm long channel with an electric field strength of 150 V/cm. As an example of the fractionation process, a target fragment of 20 bp was selected and extracted from the separation channel. The extraction was confirmed and evaluated by fluorescence imaging. High resolution and extraction fidelity were achieved by introducing new procedures for (i) extraction channel-blocking and (ii) segment transfer with cleaning. These procedures are necessary for the development of a practical, fully automated multitarget fractionation electrophoretic chip. A kind of CCD image processing method was introduced to monitor, control, and evaluate the procedure of fractionation. The resolution limits of the separation and extraction are discussed.
(Keyword)
Acrylamides / Chemical Fractionation / DNA / Electrophoresis, Microchip / Equipment Design / Fluorescence
Hideyo Ohuchi, Y Hayashibara, H Matsuda, M Onoi, M Mitsumori, M Tanaka, J Aoki, H Arai and Sumihare Noji : Diversified expression patterns of autotaxin, a gene for phospholipid-generating enzyme during mouse and chicken development, Developmental Dynamics, Vol.236, No.4, 1134-1143, 2007.
(Summary)
Autotaxin (ATX), or nucleotide pyrophosphatase-phosphodiesterase 2, is a secreted lysophospholipase D that generates bioactive phospholipids that act on G protein-coupled receptors. Here we show the expression patterns of the ATX gene in mouse and chicken embryos. ATX has a dynamic spatial and temporal expression pattern in both species and the expression domains during neural development are quite distinct from each other. Murine ATX (mATX) is expressed immediately rostral to the midbrain-hindbrain boundary, whereas chicken ATX (cATX) is expressed in the diencephalon and later in the parencephalon-synencephalon boundary. In the neural tube, cATX is expressed in the alar plate in contrast to mATX in the floor plate. ATX is also expressed in the hindbrain and various organ primordia such as face anlagen and skin appendages of the mouse and chicken. These results suggest conserved and non-conserved roles for ATX during neural development and organogenesis in these species.
Kuniko Mizuta, Satoshi Tsutsumi, Hiroshi Inoue, Yukiko Yamashita, Katsutoshi Miyatake, Katsuyuki Miyawaki, Sumihare Noji, Nobuyuki Kamata and Mitsuo Itakura : Molecular characterization of GDD1/TMEM16E, the gene product responsible for autosomal dominant gnathodiaphyseal dysplasia., Biochemical and Biophysical Research Communications, Vol.357, No.1, 126-132, 2007.
(Summary)
The human GDD1/TMEM16E gene has been found to be mutated in gnathodiaphyseal dysplasia, an unusual skeletal syndrome with autosomal dominant inheritance. The molecular and biochemical function(s) of GDD1 protein has not yet been elucidated. In this study, we examined the murine GDD1 gene expression pattern during embryonic development, and characterized the cellular and tissue localizations of its gene product using a GDD1-specific antibody. In the developing embryos, GDD1 mRNA expression was principally associated with differentiating and developing somites, with a highly complex spatiotemporal pattern that involved the myotomal and sclerotomal lineages of somites. Biochemical studies indicated that GDD1 protein is an integral membrane glycoprotein that resides predominantly in intracellular vesicles. Immunohistochemical analysis showed a high level of murine GDD1 protein expression in cardiac and skeletal muscle tissues, and in growth-plate chondrocytes and osteoblasts in bone. These observations suggest diverse cellular role(s) of GDD1 in the development of musculoskeletal system.
(Keyword)
Animals / Bone Diseases / Chromosome Disorders / Embryonic Development / Genes, Dominant / Membrane Proteins / Mice / Muscle, Skeletal / myocardium / Organ Specificity / Tissue Distribution
Taro Nakamura, Taro Mito, Yoshihisa Tanaka, Tetsuya Bando, Hideyo Ohuchi and Sumihare Noji : Involvement of the canonical Wnt/Wingless signaling in determination of the proximodistal positional values within the leg segment of the cricket Gryllus bimaculatus, Development Growth & Differentiation, Vol.49, No.2, 79-88, 2007.
Taro Mito and Sumihare Noji : Evolution of developmental systems underlying segmented body plans of bilaterian animals: insights from studies of segmentation in a cricket, Paleontological Research, Vol.10, No.4, 337-344, 2006.
(Summary)
Remarkable advances in developmental genetics in the past two decades allow us to approach the evolution of animal design by elucidating the molecular mechanisms underlying divergent body plans. The ancestry and evolution of segmented body plans in the bilaterians has been an active area of investigation in this field of study. Although segmentation mechanisms have been extensively studied for the fruit fly Drosophila melanogaster, Drosophila exhibits an evolutionarily derived mode of development, and molecular mechanisms underlying Drosophila segmentation may be unrepresentative for arthropods, even for insects. We have been studying the developmental system of the cricket, Gryllus bimaculatus, to understand more ancestral and general segmentation mechanisms for insects than those of Drosophila. In Gryllus, anterior segments are specified almost simultaneously, whereas posterior segments are specified sequentially in the extending posterior region. This mode of segmentation is general and probably ancestral for arthropods. Our RNA interference-based analyses of the functions and regulatory interactions of Gryllus orthologues of Drosophila segmentation genes have revealed surprisingly divergent aspects of the segmentation system in Gryllus in comparison with that of Drosophila. For example, the anteroposterior patterning in Gryllus is principally controlled by the caudal (cad) gene, probably without bicoid, unlike Drosophila. Comparisons of regulatory networks of segmentation genes between Gryllus and Drosophila suggest that regulatory interactions between the genes vary among insects, despite conservation of the network component genes. This implies that the molecular mechanisms of segmentation have changed dynamically during insect evolution, whereas the segmented body plan itself has been conserved. We also discuss evolution of developmental systems generating segment patterns in non-arthropod bilaterian animals.
Y Ohsawa, H Hagiwara, M Nakatani, A Yasue, K Moriyama, T Murakami, K Tsuchida, Sumihare Noji and Y Sunada : Muscular atrophy of caveolin-3-deficient mice is rescued by myostatin inhibition., The Journal of Clinical Investigation, Vol.116, No.11, 2924-2934, 2006.
K Tsuchida, Y Sunada, Sumihare Noji, T Murakami, A Uezumi and M Nakatani : Inhibitors of the TGF-β superfamily and their clinical applications., Mini Reviews in Medicinal Chemistry, Vol.6, No.11, 1255-1261, 2006.
(Summary)
The transforming growth factor-beta (TGF-beta) superfamily includes TGF-betas, activin, myostatin and bone morphogenetic proteins. Misregulation of the activity of TGF-beta family members is involved in pathogenesis of cancer, muscular dystrophy, obesity and bone and tooth remodeling. Natural inhibitors for the TGF-beta superfamily regulate fine-tuning of activity of TGF-beta family in vivo. In addition to natural inhibitors for the TGF-beta family, soluble forms of receptors for the TGF-beta family, blocking monoclonal antibodies and small chemical TGF-beta inhibitors have been developed. In this review, we summarize recent advances in our understanding of inhibitors for the TGF-beta superfamily and their medical applications.
Yohei Shinmyo, Taro Mito, T Uda, Taro Nakamura, Katsuyuki Miyawaki, Hideyo Ohuchi and Sumihare Noji : brachyenteron is necessary for morphogenesis of the posterior gut but not for anteroposterior axial elongation from the posterior growth zone in the intermediate-germband cricket Gryllus bimaculatus., Development, Vol.133, No.22, 4539-4547, 2006.
(Summary)
In the long-germband insect Drosophila, all body segments and posterior terminal structures, including the posterior gut and anal pads, are specified at the blastoderm stage. In short- and intermediate-germband insects, however, posterior segments are sequentially produced from the posterior growth zone, a process resembling somitogenesis in vertebrates, and invagination of the posterior gut starts after anteroposterior (AP) axial elongation from the growth zone. The mechanisms underlying posterior segmentation and terminal patterning in these insects are poorly understood. In order to elucidate these mechanisms, we have investigated the roles of the Brachyury/brachyenteron (Bra/byn) homolog in the intermediate-germband cricket Gryllus bimaculatus. Loss-of-function analysis by RNA interference (RNAi) revealed that Gryllus byn (Gb'byn) is not required for AP axial elongation or normal segment formation, but is required for specification of the posterior gut. We also analyzed Gryllus caudal (Gb'cad) RNAi embryos using in situ hybridization with a Gb'byn probe, and found that Gb'cad is required for internalization of the posterior gut primordium, in addition to AP axial elongation. These results suggest that the functions of byn and cad in posterior terminal patterning are highly conserved in Gryllus and Drosophila despite their divergent posterior patterning. Moreover, because it is thought that the progressive growth of the AP axis from the growth zone, controlled by a genetic program involving Cdx/cad and Bra/byn, might be ancestral to bilaterians, our data suggest that the function of Bra/byn in this process might have been lost in insects.
(Keyword)
Amino Acid Sequence / Animals / Base Sequence / Body Patterning / Cloning, Molecular / DNA Primers / DNA-Binding Proteins / Drosophila Proteins / Gryllidae / In Situ Hybridization / Lower Gastrointestinal Tract / Molecular Sequence Data / Morphogenesis / RNA Interference / Sequence Alignment / Sequence Analysis, DNA / Trans-Activators
M Tanaka, S Okudaira, Y Kishi, R Ohkawa, S Iseki, M Ota, Sumihare Noji, Y Yatomi, J Aoki and H Arai : Autotaxin stabilizes blood vessels and is required for embryonic vasculature by producing lysophosphatidic acid., The Journal of Biological Chemistry, Vol.281, No.35, 25822-25830, 2006.
Masahiro Shin, Sumihare Noji, Annette Neubuser and Sadao Yasugi : FGF10 is required for cell proliferation and gland formation in the stomach epithelium of the chicken embryo, Developmental Biology, Vol.294, No.1, 11-23, 2006.
(Summary)
The development of digestive organs in vertebrates involves active epithelial-mesenchymal interactions. In the chicken proventriculus (glandular stomach), the morphogenesis and cytodifferentiation of the epithelium are controlled by the inductive signaling factors that are secreted from the underlying mesenchyme. Previous studies have shown that Fgf10 is expressed in the developing chicken proventricular mesenchyme, whereas its receptors are present in the epithelium. In our present study, we show that FGF10 is an early mesenchymal signal that is critically associated with the developmental processes in the proventricular epithelium. Furthermore, virus-mediated Fgf10 overexpression in ovo results in a hypermorphic epithelial structure and an increase in epithelial cell number. In contrast, the overexpression of a secreted FGFR2b (sFGFR2b), an FGF10 antagonist, blocks cell proliferation and gland formation in the proventricular epithelium in ovo. This downregulation of proliferative activity was subsequently found to retard gland formation and also to delay differentiation of the epithelium. These results demonstrate that FGF10 signaling, mediated by FGFR1b and/or FGFR2b, is required for proliferation and gland formation in the epithelium in the developing chick embryo.
Hirotaka Tao, K Ono, Hitomi Kurose, Sumihare Noji and Hideyo Ohuchi : Exogenous FGF10 can rescue an eye-open at birth phenotype of Fgf10-null mice by activating activin and TGFalpha-EGFR signaling., Development Growth & Differentiation, Vol.48, No.5, 339-346, 2006.
N Wada, Tsutomu Nohno and Sumihare Noji : [Roles of the BMP family in pattern formation of the vertebrate limb], Clinical Calcium, Vol.16, No.5, 773-780, 2006.
(Summary)
In the process of limb development, various signaling molecules are produced in the organizing center of the limb bud. These molecules regulate pattern formation and the morphogenesis of the limb. Members of the bone morphogenetic protein (BMP) family and their receptors are expressed in the limb bud in spatiotemporal specific patterns. BMP signaling regulates chondrogenesis of limb mesenchyme, and promotes apoptosis of the interdigital soft tissue. Recently, this signaling was found to be involved in establishment of dorsoventral polarity of the limb bud and in regulation of limb growth via maintenance of the apical ectodermal ridge. BMPs also regulate digit morphology. In these events, BMPs work together with other signaling molecules, but they sometimes act as negative regulators of these other molecules so that limb morphogenesis can proceed normally.
A Yoshimura, A Nakata, Taro Mito and Sumihare Noji : The characteristics of karyotype and telomeric satellite DNA sequences in the cricket, Gryllus bimaculatus (Orthoptera, Gryllidae), Cytogenetic and Genome Research, Vol.112, No.3-4, 329-336, 2006.
(Summary)
The chromosomes derived from the Japanese population of Gryllus bimaculatus were characterized by C-banding and Ag-NOR staining. The chromosome number, 2n = 28 + XX (female)/XO (male), corresponded with that of other populations of G. bimaculatus, but the chromosome configuration in idiograms varied between the populations. NORs were carried on one pair of autosomes and appeared polymorphous. The positive C-bands located at the centromere of all chromosomes and the distal regions of many chromosome pairs, and the size and the distribution pattern of the distal C-heterochromatin showed differences among the chromosomes. In addition, this paper reports on the characteristics of HindIII satellite DNA isolated from the genome of G. bimaculatus. The HindIII repetitive fragments were about 0.54 kb long, and localized at the distal C-bands of the autosomes and the interstitial C-bands of the X chromosome. Molecular analysis showed two distinct satellite DNA sequences, named the GBH535 and GBH542 families, with high AT contents of about 67 and 66%, respectively. The two repetitive families seem to be derived from a common ancestral sequence, and both families possessed the same 13-bp palindrome sequence. The results of Southern blot hybridization suggest that the sequence of the GBH535 family is conserved in the genomic DNAs of Gryllus species, whereas the GBH542 family is a species-specific sequence.
(Keyword)
Animals / Antigens, Nuclear / Chromosome Banding / Chromosome Mapping / DNA, Satellite / Female / Gryllidae / Japan / Karyotyping / Male / Nuclear Proteins / Telomere / X Chromosome / Y Chromosome
Sayuri Tomonari, Shino Akamatsu, Sumihare Noji and Hideyo Ohuchi : A non-canonical photopigment, melanopsin, is expressed in the differentiating ganglion, horizontal, and bipolar cells of the chicken retina, Developmental Dynamics, Vol.234, No.3, 783-790, 2005.
(Summary)
Vertebrate melanopsin is a photopigment in the eye, required for photoentrainment. Melanopsin is more closely related to opsin proteins found in invertebrates, than to the other photo-pigments. Although the invertebrate melanopsin-like protein is localized in rhabdomeric photoreceptors in the invertebrate eye, it has been shown to be expressed in a subset of retinal ganglion cells in the mouse and in horizontal cells in the frog, indicating its diversified expression pattern in vertebrates. Here we show that two types of melanopsin transcripts are expressed in the developing chicken retina. Melanopsin is firstly expressed by a small subset of ganglion cells, and then prominently expressed by horizontal cells and later by bipolar cells in the developing chicken retina. This suggests that a subset of ganglion, horizontal, and bipolar cells in the chicken retina may have rhabdomeric properties in their origins.
Michio Ogasawara, Nori Satoh, Yasuhito Shimada, Zhipeng Wang, Toshio Tanaka and Sumihare Noji : Rapid and stable buffer exchange system using InSitu Chip suitable for multicolor and large-scale whole-mount analyses., Development Genes and Evolution, Vol.216, No.2, 100-104, 2005.
(Summary)
Whole-mount in situ hybridization (WISH) and whole-mount immunohistochemistry (WIHC) are informative methods commonly used to analyze the spatiotemporal and quantitative distribution of mRNAs and proteins. However, these methods require multiple buffer changes and the imposition of time- and nerve-consuming efforts. To facilitate the whole-mount analyses, we innovated an easy and one-step buffer exchange system named "InSitu Chip" based on a single column containing two attached filters. This system improves the speed and stabilizes the different steps of the currently available protocols, providing fast and uniform operations. The InSitu Chip system is especially appropriate for multicolor whole-mount analyses using fluorescent detection. Furthermore, the InSitu Chip system is also suitable for large-scale whole-mount experiments associated with genome, transcriptome, and/or proteome analyses requiring high-throughput, high-quality, and reproducible results. Using the InSitu Chip, about 1,500 gene expression patterns were stably surveyed in ascidian Ciona intestinalis juveniles.
(Keyword)
Whole / mount analyses / WISH / WIHC / High throughput / InSitu Chip
Hirotaka Tao, Miyuki Shimizu, Ryo Kusumoto, Katsuhiko Ono, Sumihare Noji and Hideyo Ohuchi : A dual role of FGF10 in proliferation and coordinated migration of epithelial leading edge cells during mouse eyelid development., Development, Vol.132, No.14, 3217-3230, 2005.
(Summary)
The development of the eyelid requires coordinated cellular processes of proliferation, cell shape changes, migration and cell death. Mutant mice deficient in the fibroblast growth factor 10 (Fgf10) gene exhibit open-eyelids at birth. To elucidate the roles of FGF10 during eyelid formation, we examined the expression pattern of Fgf10 during eyelid formation and the phenotype of Fgf10-null eyelids in detail. Fgf10 is expressed by mesenchymal cells just beneath the protruding epidermal cells of the nascent eyelid. However, Fgf10-null epithelial cells running though the eyelid groove do not exhibit typical cuboid shape or sufficient proliferation. Furthermore, peridermal clumps are not maintained on the eyelid leading edge, and epithelial extension does not occur. At the cellular level, the accumulation of actin fibers is not observed in the mutant epithelial leading edge. The expression of activin/inhibin betaB (ActbetaB/Inhbb) and transforming growth factor alpha (Tgfa), previously reported to be crucial for eyelid development, is down-regulated in the mutant leading edge, while the onset of sonic hedgehog (Shh) expression is delayed on the mutant eyelid margin. Explant cultures of mouse eyelid primordia shows that the open-eyelid phenotype of the mutant is reduced by exogenous FGF10 protein, and that the expression of ActbetaB and Tgfa is ectopically induced in the thickened eyelid epithelium by the FGF10 protein. These results indicate a dual role of FGF10 in mouse eyelid development, for both proliferation and coordinated migration of eyelid epithelial cells by reorganization of the cytoskeleton, through the regulation of activin, TGFalpha and SHH signaling.
Hideyo Ohuchi, Akihiro Yasue, Katsuhiko Ono, Shunsuke Sasaoka, Sayuri Tomonari, Akira Takagi, Mitsuo Itakura, Keiji Moriyama, Sumihare Noji and Tsutomu Nohno : Identification of Cis-Element Regulating Expression of the Mouse Fgf10 Gene during Inner Ear Development, Developmental Dynamics, Vol.233, No.1, 177-187, 2005.
(Summary)
Fibroblast growth factor (FGF) signaling is crucial for the induction and growth of the ear, a sensory organ that involves intimate tissue interactions. Here, we report the abnormality of Fgf10 null ear and the identification of a cis-regulatory element directing otic expression of Fgf10. In Fgf10 null inner ears, we found that the initial development of semicircular, vestibular, and cochlear divisions is roughly normal, after which there are abnormalities of semicircular canal/cristae and vestibular development. The mutant semicircular disks remain without canal formation by the perinatal stage. To elucidate regulation of the Fgf10 expression during inner ear development, we isolated a 6.6-kb fragment of its 5'-upstream region and examined its transcriptional activity with transgenic mice, using a lacZ-reporter system. From comparison of the mouse sequences of the 6.6-kb fragment with corresponding sequences of the human and chicken Fgf10, we identified a 0.4-kb enhancer sequence that drives Fgf10 expression in the developing inner ear. The enhancer sequences have motifs for many homeodomain-containing proteins (e.g., Prx, Hox, Nkx), in addition to POU-domain factors (e.g., Brn3), zinc-finger transcription factors (e.g., GATA-binding factors), TCF/LEF-1, and a SMAD-interacting protein. Thus, FGF10 signaling is dispensable for specification of otic compartment identity but is required for hollowing the semicircular disk. Furthermore, the analysis of a putative inner ear enhancer of Fgf10 has disclosed a complicated regulation of Fgf10 during inner ear development by numerous transcription factors and signaling pathways.
Isao Sarashina, Taro Mito, Michiko Saito, Hiroyuki Uneme, Katsuyuki Miyawaki, Yohei Shinmyo, Hideyo Ohuchi and Sumihare Noji : Location of micropyles and early embryonic development of the two-spotted cricket Gryllus bimaculatus (Insecta, Orthoptera), Development Growth & Differentiation, Vol.47, No.2, 99-108, 2005.
(Summary)
Early embryogenesis of the two-spotted cricket Gryllus bimaculatus was examined by scanning electron microscopy and several fluorescence staining methods, with special reference to these four issues: (i) the location of micropyles; (ii) the transfer of the female pronucleus following meiosis; (iii) the timing of cellularization; and (iv) the process of the germ primordium formation. Between two and four micropyles lie in the mid-ventral region of the egg. The egg nucleus is at the mid-dorsal periphery of the new laid egg, and meiosis resumes and is completed there. The female pronucleus moves to the mid-ventral side, and fertilization occurs there. Energid starts to proliferate and migrates to the periphery of the egg, initiating blastoderm formation. Actin caps surround each superficial nucleus. Cellularization occurs during the blastoderm stage. At a late blastoderm stage, nuclei aggregate in both the posterolateral patch-like regions of the egg to form a germ primordium. The germ primordium looks like a pair of dumbbells. Both the patches shift towards the ventral side and fuse into a germ primordium. The germ primordium contracts to produce a clearly delineated germ band. Observations on distribution patterns of F-actin indicate that, all through the process, the germ primordium retains that unity, and is not separated into two parts.
Yohei Shinmyo, Taro Mito, Taro Matsushita, Isao Sarashina, Katsuyuki Miyawaki, Hideyo Ohuchi and Sumihare Noji : caudal is required for gnathal and thoracic patterning and for posterior elongation in the intermediate-germband cricket Gryllus bimaculatus, Mechanisms of Development, Vol.122, No.2, 231-239, 2005.
(Summary)
Although the molecular mechanisms directing anteroposterior patterning of the Drosophila embryo (long-germband mode) are well understood, how these mechanisms were evolved from an ancestral mode of insect embryogenesis remains largely unknown. In order to gain insight into mechanisms of evolution in insect embryogenesis, we have examined the expression and function of the orthologue of Drosophila caudal (cad) in the intermediate-germband cricket Gryllus bimaculatus. We observed that a posterior (high) to anterior (low) gradient in the levels of Gryllus bimaculatus cad (Gb' cad) transcript was formed in the early-stage embryo, and then Gb' cad was expressed in the posterior growth zone until the posterior segmentation was completed. Reduction of Gb' cad expression level by RNA interference resulted in deletion of the gnathum, thorax, and abdomen in embryos, remaining only anterior head. We found that the gnathal and thoracic segments are formed by Gb' cad probably through the transcriptional regulation of gap genes including Gb' hunchback and Gb' Kruppel. Furthermore, Gb' cad was found to be involved in the posterior elongation, acting as a downstream gene in the Wingless/Armadillo signalling pathways. These findings indicate that Gb' cad does not function as it does in Drosophila, suggesting that regulatory and functional changes of cad occurred during insect evolution. Since Wnt/Cdx pathways are involved in the posterior patterning of vertebrates, such mechanisms may be conserved in animals that undergo sequential segmentation from the posterior growth zone.
Hiroyuki Iwahana, Morisada Hayakawa, Kenji Kuroiwa, Kenji Tago, Ken Yanagisawa, Sumihare Noji and S Tominaga : Molecular cloning of the chicken ST2 gene and a novel variant form of the ST2 gene product, ST2LV, Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression, Vol.1681, No.1, 1-14, 2004.
Katsuyuki Miyawaki, Taro Mito, Isao Sarashina, Hongjie Zhang, Yohei Shinmyo, Hideyo Ohuchi and Sumihare Noji : Involvement of Wingless/Armadillo signaling in the posterior sequential segmentation in the cricket, Gryllus bimaculatus (Orthoptera), as revealed by RNAi analysis, Mechanisms of Development, Vol.121, No.2, 119-130, 2004.
F Myokai, E Koyama, K Nishikawa, Sumihare Noji, Y Murayama and S Taniguchi : Aspects of interleukin-8 gene expression by gingival and dermal fibroblasts stimulated with interleukin-1beta or tumour necrosis factor alpha, Journal of the International Academy of Periodontology, Vol.6, No.1, 21-28, 2004.
Yukihisa Matsumoto, Sumihare Noji and Makoto Mizunami : Time Course of Protein Synthesis-Dependent Phase of Olfactory Memory in the Cricket Gryllus bimaculatus, Zoological Science, Vol.20, No.4, 409-416, 2003.
Taizo Horikoshi, Naoto Endo, Minoru Shibata, Peter Heutink, Rovert E. Hill and Sumihare Noji : Disruption of the C7orf2/Lmbr1 genic region is associated with preaxial polydactyly in humans and mice, Journal of Bone and Mineral Metabolism, Vol.21, No.1, 1-4, 2003.
Hideyo Ohuchi, Hirotaka Tao, Kazuyo Ohata, Nobuyuki Itoh, Shigeaki Kato and Sumihare Noji : Fibroblast growth factor 10 is required for proper development of the mouse whiskers, Biochemical and Biophysical Research Communications, Vol.302, No.3, 562-567, 2003.
Hongjie Zhang, Yohei Shinmyo, Ayumi Hirose, Taro Mito, Yoshiko Inoue, Hideyo Ohuchi, Thanasis G. Loukeris, Paul Eggleston and Sumihare Noji : Extrachromosomal transposition of the transposable element Minos in embryos of the cricket Gryllus bimaculatus, Development Growth & Differentiation, Vol.44, No.5, 409-417, 2002.
(Summary)
Effective germline transformation of insects has been shown to depend on the right choice of transposon system and selection marker. In this study the promoter region of a Gryllus cytoplasmic actin (GbA3/4) gene was isolated and characterized, and was used to drive the expression of Minos transposase in embryos of the cricket Gryllus bimaculatus. Active Minos transposase was produced in these embryos as monitored through established transposon excision and interplasmid transposition assays. In contrast, Drosophila melanogaster hsp70 promoter, previously used to express Minos transposase in a number of insect species and insect cell lines, failed to produce any detectable Minos transposase activity, as recorded by using the very sensitive transposon excision assay. In addition, the GbA3/4 promoter was found to drive expression of enhanced green fluorescent protein (eGFP) predominantly in vitellophages of the developing Gryllus eggs when a plasmid carrying a GbA3/4 promoter-eGFP fusion gene was transiently injected into embryos. These results strongly support the use of Minos transposons marked with the GbA3/4 promoter-eGFP for the genetic transformation of this emerging model insect species.
(Keyword)
Actins / Amino Acid Sequence / Animals / Base Sequence / DNA Transposable Elements / Genes, Reporter / Genetic Vectors / Gryllidae / Molecular Sequence Data / Plasmids / Promoter Regions, Genetic
Hirotaka Tao, Yasuko Yoshimoto, Hidefumi Yoshioka, Tsutomu Nohno, Sumihare Noji and Hideyo Ohuchi : FGF10 is a mesenchymally derived stimulator for epidermal development in the chick embryonic skin, Mechanisms of Development, Vol.116, No.1-2, 39-49, 2002.
Taro Mito, Yoshiko Inoue, Shinsuke Kimura, Katsuyuki Miyawaki, Nao Niwa, Yohei Shinmyo, Hideyo Ohuchi and Sumihare Noji : Involvement of hedgehog, wingless, and dpp in the initiation of proximodistal axis formation during the regeneration of insect legs, a verification of the modified boundary model, Mechanisms of Development, Vol.114, No.1, 27-35, 2002.
Yoshiko Inoue, Taro Mito, Katsuyuki Miyawaki, Kyoko Matsushima, Yohei Shinmyo, Tiffany A. Heanue, Graeme Mardon, Hideyo Ohuchi and Sumihare Noji : Correlation of expression patterns of homothorax, dachshund, and Distal-less with the proximodistal segmentation of the cricket leg bud, Mechanisms of Development, Vol.113, No.2, 141-148, 2002.
Laura A. Lettice, Taizo Horikoshi, Simon J.H. Heaney, Marijke J. van Baren, C. Herma Linde, J. Guido Breedveld, Marijke Joosse, Nurten Akarsu, Ben A. Oostra, Naoto Endo, Minoru Shibata, Mikio Suzuki, Eiichi Takahashi, Toshikatsu Shinka, Yutaka Nakahori, Dai Ayusawa, Kazuhiko Nakabayashi, Stephen W. Scherer, Peter Heutink, Robert E. Hill and Sumihare Noji : Disruption of a long-range cis-acting regulator for Shh causes preaxial polydactyly, Proceedings of the National Academy of Sciences of the United States of America, Vol.99, No.11, 7548-7553, 2002.
(Summary)
Preaxial polydactyly (PPD) is a common limb malformation in human. A number of polydactylous mouse mutants indicate that misexpression of Shh is a common requirement for generating extra digits. Here we identify a translocation breakpoint in a PPD patient and a transgenic insertion site in the polydactylous mouse mutant sasquatch (Ssq). The genetic lesions in both lie within the same respective intron of the LMBR1/Lmbr1 gene, which resides approximately 1 Mb away from Shh. Genetic analysis of Ssq reveals that the Lmbr1 gene is incidental to the phenotype and that the mutation directly interrupts a cis-acting regulator of Shh. This regulator is most likely the target for generating PPD mutations in human.
Takashi Yamaoka, Kenji Yoshino, Taketo Yamada, Mikiko Yano, Takefumi Matsui, Takashi Yamaguchi, Maki Moritani, Jun-ichi Hata, Sumihare Noji and Mitsuo Itakura : Transgenic expression of FGF8 and FGF10 induces transdifferentiation of pancreatic islet cells into hepatocytes and exocrine cells, Biochemical and Biophysical Research Communications, Vol.292, No.1, 138-143, 2002.
(Summary)
FGF signaling is essential for normal development of pancreatic islets. To examine the effects of overexpressed FGF8 and FGF10 on pancreatic development, we generated FGF8- and FGF10-transgenic mice (Tg mice) under the control of the glucagon promoter. In FGF8-Tg mice, hepatocyte-like cells were observed in the periphery of pancreatic islets, but areas of alpha and beta cells did not decrease, whereas in FGF10-Tg mice, pancreatic ductal and acinar cells were found in islets, concomitantly with disturbed beta-cell differentiation. These results suggest that FGF8 and FGF10 play important roles in development of hepatocytes and exocrine cells, respectively, and explain the absence of FGF8 expression in normal islets and pancreatic hypoplasia in FGF10-deficient mice.
Yoshiko Inoue, Nao Niwa, Taro Mito, Hideyo Ohuchi, Hidefumi Yoshioka and Sumihare Noji : Expression patterns of hedgehog, wingless, and decapentaplegic during gut formation of Gryllus bimaculatus (cricket), Mechanisms of Development, Vol.110, No.1-2, 245-248, 2002.
Hidemi Sasaki, Takashi Yamaoka, Hideyo Ohuchi, Akihiro Yasue, Tsutomu Nohno, Hirotaka Kawano, Shigeaki Kato, Mitsuo Itakura, Masaru Nagayama and Sumihare Noji : Identification of cis-elements regulating expression of Fgf10 during limb development., The International Journal of Developmental Biology, Vol.46, No.7, 963-967, 2002.
(Summary)
Fibroblast growth factor 10 (FGF10) is known to be expressed in limb mesenchymal cells and to function as a mesenchymal signaling factor involved in epithelial-mesenchymal interactions during limb development. To elucidate regulation of Fgf10 expression, we isolated the promoter region of Fgf10 containing its 2.0 kb upstream 5'-fragment from the initiation codon and its 0.9 kb downstream fragment. Transcriptional activity of the fragment was examined with transgenic mice, using a lacZ-reporter system. Although no significant expression of the reporter gene was observed for the 0.2 kb 5'-fragment, expression was detected in the apical ectodermal ridge of the limb bud and developing cartilage of the limb for the 2.0 kb and 0.7 kb 5'-fragments, respectively. From comparison of the mouse sequences of the 2.0 kb fragment with corresponding sequences of human and chicken Fgf10, we identified 17 conserved putative enhancer motifs for AER expression and other unidentified expressions. For limb cartilage expression, we found putative enhancer sequences conserved among the three species in the 0.7 kb 5'-fragment. In the fragment, three DNA binding motifs were identified in the mouse and human sequence, although they are not conserved in the corresponding chicken sequence.
Nobuki Matuura, Chichung D. Lie, Minoru Hoshimaru, Minoru Asahi, Masato Hojo, Ryuji Ishizaki, Nobuo Hashimoto, Sumihare Noji, Hideyo Ohuchi, Hidefumi Yoshioka and H. Fred Gage : Sonic hedgehog facilitates dopamine differentiation in the presence of a mesencephalic glial cell line, The Journal of Neuroscience, Vol.21, No.12, 4326-4335, 2001.
Akihiro Ohyama, Fumi Saito, Hideyo Ohuchi and Sumihare Noji : Differential expression of two BMP antagonists, gremlin and Follistatin, during development of the chick feather bud, Mechanisms of Development, Vol.100, No.2, 331-333, 2001.
(Summary)
Expression of four BMP antagonist genes, noggin, chordin, gremlin and Follistatin, was examined during chick feather development. Although expression of noggin and chordin was not detected, gremlin and Follistatin were expressed differentially in feather buds. The differential expression patterns of gremlin and Follistatin change dynamically from the nascent inter-feather bud region to the posterior domain of the feather bud.
(Keyword)
Animals / Chick Embryo / Feathers / Follistatin / Gene Expression / Glycoproteins / In Situ Hybridization / Mesoderm / Protein Biosynthesis / Time Factors
Akihiro Yasue, Hirotaka Tao, Tsutomu Nohno, Keiji Moriyama, Sumihare Noji and Hideyo Ohuchi : Cloning and expression of the chick p63 gene, Mechanisms of Development, Vol.100, No.1, 105-108, 2001.
(Summary)
We have isolated a chick cDNA for p63, a member of the p53 transcription factor family. This cDNA encodes a protein of 582 amino acids for an alpha isoform in the C-terminal region, while lacking the N-terminal transactivation domain. The chick p63 gene is first expressed in the prospective cutaneous ectoderm at stage 6 and later in the developing epithelia. The p63 expression is intense in specialized epithelial structures, such as apical ectodermal ridge of the limb bud, epithelia of branchial arches and feather buds. Furthermore, we have found that the transcripts are detected in the interdigital epithelium, intersomite epithelium, and epaxial dermamyotome.
Nao Niwa, Yoshiko Inoue, Akiyoshi Nozawa, Mariko Saito, Yoshio Misumi, Hideyo Ohuchi, Hidefumi Yoshioka and Sumihare Noji : Correlation of diversity of leg morphology in gryllus bimaculatus (cricket) with divergence in dpp expression pattern during leg development, Development, Vol.127, No.20, 4373-4381, 2000.
Motomi Enomoto-Iwamoto, Takashi Nakamura, Tomonao Aikawa, Yoshinobu Higuchi, Takahito Yuasa, Akira Yamaguchi, Tsutomu Nohno, Sumihare Noji, Tokuzo Matsuya, Kojiro Kurisu, Masahiro Iwamoto, Maurizio Pacifici and Eiki Koyama : Hedgehog proteins stimulate chondrogenic cell differentiation and cartilage formation, Journal of Bone and Mineral Research, Vol.15, No.9, 1659-1668, 2000.
116.
Jun Kimura, Mika Sato-Maeda, Sumihare Noji and Hiroyuki Ide : Synergistic effects of FGF and non-ridge ectoderm on gene expression involved in the formation of the anteroposterior axis of the chick limb bud in cell culture, Development Growth & Differentiation, Vol.42, No.3, 219-227, 2000.
Yasuhiko Kawakami, Naoyuki Wada, Shin-ichiro Nishimatsu, Chikako Komaguchi, Sumihare Noji and Tsutomu Nohno : Identification of chick frizzled-10 expressed in the developing limb and the central nervous system, Mechanisms of Development, Vol.91, No.1-2, 375-378, 2000.
(Summary)
We have identified chick frizzled (Fz)-10, encoding a Wnt receptor, and examined the expression pattern during embryogenesis. Fz-10 is expressed in the region posterior to the Hensen's node at stage 6. Fz-10 expression is detected in the dorsal domain of the neural tube and the central nervous system of the developing embryo. In the developing limb, Fz-10 expression starts at stage 18 in the posterior-dorsal region of the distal mesenchyme, and gradually expands to the anterior-distal region. Fz-10 is also expressed in the feather bud and branchial arch. Implantation of Sonic hedgehog (Shh)-expressing cells into the anterior margin of the limb bud resulted in the induction of Fz-10 expression in anterior-dorsal mesenchyme.
Yoshiyasu Ishimaru, Hidefumi Yoshioka, Hirotaka Tao, Bernard Thisse, Christine Thisse, Wright V E Christopher, Hiroshi Hamada, Hideyo Ohuchi and Sumihare Noji : Asymmetric expression of antivin/lefty1 in the early chick embryo, Mechanisms of Development, Vol.90, No.1, 115-118, 2000.
(Summary)
Mammalian lefty and zebrafish antivin, highly related to lefty, are shown to be expressed asymmetrically and involved in the specification of the left body side of early embryos. We isolated a chick homologue of the antivin/lefty1 cDNA and studied its expression pattern during early chick development. We found that antivin/lefty1 is expressed asymmetrically on the left side of the prospective floorplate, notochord and lateral plate mesoderm of the chick embryo.
Gail Lizarraga, Deborah Ferrari, Michael Kalinowski, Hideyo Ohuchi, Sumihare Noji, RobertA Kosher and CarolineN Dealy : FGFR2 signaling in normal and limbless chick limb buds, Developmental Genetics, Vol.25, No.4, 331-338, 1999.
(Summary)
FGF10 and FGF8, which are reciprocally expressed by the mesoderm and AER of the developing limb bud, have been implicated in limb initiation, outgrowth, and patterning. FGF10 and FGF8 signal through the FGFR2b and FGFR2c alternative splice isoforms, respectively [Ornitz DM, et al. 1996. J Biol Chem 271:15292-15297; Igarashi M, et al. 1998. J Biol Chem 273:13230-13235]. A paracrine signaling loop model has been proposed whereby FGF10 expressed by limb mesoderm signals via ectodermally restricted FGFR2b to regulate FGF8 expression by the apical ectoderm; in turn, FGF8 signals via mesodermally restricted FGFR2c to maintain FGF10 expression [Ohuchi H, et al. 1997. Development 124:2235-2244; Xu X, et al. 1998. Development 125:753-765]. To explore this model, we have examined FGFR2b and FGFR2c mRNA expression, using isoform-specific probes during the early stages of development of the chick limb when limb initiation, AER induction, and outgrowth are occurring. We have found that FGFR2b is expressed by limb ectoderm, including the AER, consistent with paracrine signaling of FGF10. By contrast, FGFR2c is expressed by both mesoderm and ectoderm, indicating that FGF8 has the potential to function in an autocrine as well as paracrine fashion. Indeed, as the limb grows out in response to the AER, FGFR2c expression attenuates in the mesoderm of the progress zone, but is maintained in the AER itself, arguing against exclusive paracrine signaling of FGF8 during limb outgrowth. We also report that transcripts for FGF10, FGFR2b, and FGFR2c are expressed normally in the limb buds of limbless mutant embryos, which fail to form an AER and do not express FGF8. Furthermore, we detect no mutations in exons specific for the FGFR2c or FGFR2b isoforms in limbless embryos. Since gene targeting has shown that expression of FGF8 in limb ectoderm depends on FGF10 [Min H, et al. 1998. Genes Dev 12:3156-3161; Sekine K, et al. 1999. Nature Genet 21:138-141], these results indicate that the product of the limbless gene is required for FGF10 to induce expression of FGF8.
Hideyo Ohuchi, Takashi Nakagawa, Nobuyuki Itoh and Sumihare Noji : FGF10 can induce Fgf8 expression concomitantly with En1 and R-fng expression in chick limb ectoderm, independent of its dorsoventral specification, Development Growth & Differentiation, Vol.41, No.6, 665-673, 1999.
Hideyo Ohuchi, Sayuri Tomonari, Hiroyuki Itoh, Takashi Mikawa and Sumihare Noji : Identification of chick rax/rx genes with overlapping patterns of expression during early eye and brain development, Mechanisms of Development, Vol.85, No.1-2, 193-195, 1999.
122.
Hideyo Ohuchi and Sumihare Noji : Fibroblast-growth-factor-induced additional limbs in the study of initiation of limb formation, limb identity, myogenesis, and innervation, Cell and Tissue Research, Vol.296, No.1, 45-56, 1999.
(Summary)
In this review, we focus on the additional limb induced by members of the fibroblast growth factor (FGF) family in the flank of chick embryos. The "additional limb" was first reported 73 years ago by Balinsky in 1925. He grafted otic vesicle to the flank of newt embryos and observed the formation of the "additional limb." In 1995, formation of an additional limb was found to be induced by FGF in the chick embryo. This finding subsequently led to the recent understanding of how the limb bud is initially formed, how the limb position is determined, and how the limb identity is determined. Thus, the additional limb has been recognized as a useful experimental system for the study of limb development and its relation to the regionalization of the body. Furthermore, since limb muscles are formed from cells which have migrated from somites and innervation to them takes place from the spinal cord, the additional limb would also be a powerful tool with which to study the relation of limb morphogenesis to developmental processes of the spinal cord and somites. This review consists of five sections: (1) "Introduction," (2) "How to make additional limbs," (3) "Characteristics of the additional limb," (4) "Studies with the additional limb," and (5) "Concluding remarks." In the second section, techniques to make additional limbs are reviewed, showing that additional limbs can be made by fairly easy manipulation of the chick embryo. In the third section, the characteristics analyzed so far of the additional limb are summarized, focusing on its morphology. In the fourth section, recent studies on the use of the additional limb are reviewed: experiments on the additional limb have been performed to elucidate the mechanisms governing determination of limb identity by Hox codes and the Tbx family and initiation of limb formation by FGF10. In addition, the roles of SF/HGF in the formation of limb muscles have also been investigated using the additional limb. In the near future, the additional limb will be also used in the study of innervation from the spinal cord, and probably migration of neural crest cells.
Yasuhiko Kawakami, Naoyuki Wada, Shin-ichiro Nishimatsu, Tetsuya Ishikawa, Sumihare Noji and Tsutomu Nohno : Involvement of Wnt-5a in chondrogenic pattern formation in the chick limb bud, Development Growth & Differentiation, Vol.41, No.1, 29-40, 1999.
124.
Jun-ichi Funahashi, Tatsuya Okafuji, Hideyo Ohuchi, Sumihare Noji, Hideaki Tanaka and Harukazu Nakamura : Role of Pax-5 in the regulation of a mid-hindbrain organizer's activity, Development Growth & Differentiation, Vol.41, No.1, 59-72, 1999.
(Summary)
The mes-metencephalic boundary (isthmus) has been suggested to act as an organizer in the development of the optic tectum. Pax-5 was cloned as a candidate for regulator of the organizing center. Isthmus-specific expression of Pax-5 and analogy with the genetic cascade in Drosophila suggest that Pax-5 may be at a higher hierarchical position in the gene regulation cascade of tectum development. To examine this possibility, a gain-of-function experiment on Pax-5 was carried out. In ovo electroporation on E2 chick brain with the eucaryotic expression vector that encodes chick Pax-5 cDNA was used. Not only was a considerable amount of Pax-5 expressed ectopically in the transfected brain, but irregular bulging of the neuroepithelium was induced in the diencephalon and mesencephalon. At Pax-5 misexpressing sites, uptake of BrdU was increased. Histological examination of E7 transfected brain revealed that Pax-5 caused transdifferentiation of diencephalon into the tectum-like structure. In the bulges of the E7 mesencephalon, differentiation of laminar structure was repressed when compared to the normal side. In transfected embryos, En-2, Wnt-1 and Fgf8 were up-regulated ectopically, and Otx2 was down-regulated in the diencephalon to mesencephalon. Moreover, Ephrin-A2, which is expressed specifically in the tectum with a gradient highest at the caudal end, is suggested to be involved in pathfinding of the retinal fibers, and was induced in the bulges. When the mouse Fgf8 expression vector was electroporated, Pax-5 and chick Fgf8 were also induced ectopically. These results suggest that Pax-5, together with Fgf8, hold a higher position in the genetic hierarchy of the isthmus organizing center and regulate its activity.
G Lizarraga, D Ferrari, M kalinowski, Hideyo Ohuchi, Sumihare Noji, R.A Kosher and C.N Dealy : Isoform-specific FGFR2 expression during chick limb development and use of the amelic limbless mutant to explore FGF10/FGF8 signaling, Developmental Genetics, Vol.25, 331-338, 1999.
126.
Chikara Meno, Akihiko Shimono, Yukio Saijoh, Kenta Yashiro, Kyoko Mochida, Sachiko Ohishi, Sumihare Noji, Hisato Kondoh and Hiroshi Hamada : lefty-1 is required for left-right determination as a regulator of lefty-2 and nodal, Cell, Vol.94, No.3, 287-297, 1998.
(Summary)
lefty-1, lefty-2, and nodal are expressed on the left side of developing mouse embryos and are implicated in left-right (L-R) determination. The role of lefty-1 was examined by analyzing mutant mice lacking this gene. The lefty-1-deficient mice showed a variety of L-R positional defects in visceral organs. Unexpectedly, however, the most common feature of lefty-1-/- mice was thoracic left isomerism (rather than right isomerism). The lack of lefty-1 resulted in bilateral expression of nodal, lefty-2, and Pitx2 (a homeobox gene normally expressed on the left side). These observations suggest that the role of lefty-1 is to restrict the expression of lefty-2 and nodal to the left side, and that lefty-2 or nodal encodes a signal for "leftness."
Hidefumi Yoshioka, Chikara Meno, Kazuko Koshiba, Minoru Sugihara, Hiroyuki Itoh, Yoshiyasu Ishimaru, Takashi Inoue, Hideyo Ohuchi, Elena V Semina, Jeffrey C Murray, Sumihare Noji and Hiroshi Hamada : Pitx2, a bicoid-type homeobox gene, is involved in a lefty-signaling pathway in determination of left-right asymmetry, Cell, Vol.94, No.3, 299-305, 1998.
(Summary)
Signaling molecules such as Activin, Sonic hedgehog, Nodal, Lefty, and Vg1 have been found to be involved in determination of left-right (L-R) asymmetry in the chick, mouse, or frog. However, a common signaling pathway has not yet been identified in vertebrates. We report that Pitx2, a bicoid-type homeobox gene expressed asymmetrically in the left lateral plate mesoderm, may be involved in determination of L-R asymmetry in both mouse and chick. Since Pitx2 appears to be downstream of lefty-1 in the mouse pathway, we examined whether mouse Lefty proteins could affect the expression of Pitx2 in the chick. Our results indicate that a common pathway from lefty-1 to Pitx2 likely exists for determination of L-R asymmetry in vertebrates.
Takashi Yamaoka, Chiyoko Idehara, Makiko Yano, Takaya Matsushita, Taketo Yamada, Setsuko Ii, Maki Moritani, Jun-ichi Hata, Hiromu Sugino, Sumihare Noji and Mitsuo Itakura : Hypoplasia of pancreatic islets in transgenic mice expressing activin receptor mutants, The Journal of Clinical Investigation, Vol.102, No.2, 294-301, 1998.
(Summary)
Activin, a member of the TGF-beta superfamily, regulates the growth and differentiation of a variety of cell types. Based on the expression of activin in pancreatic rudiments of rat embryos and stimulation of insulin secretion from adult rat pancreatic islets by activin, activin is implicated in the development and function of islets. To examine the significance of activin signaling in the fetal and postnatal development of islets, transgenic mice expressing a dominant negative form of activin receptor (dn-ActR) or a constitutively active form of activin receptor (ActR-T206D) in islets were generated together with the transgenic mice expressing intact activin receptor (intact ActR) as a negative control. Transgenic mice with both dn-ActR and ActR-T206D showed lower survival rates, smaller islet area, and lower insulin content in the whole pancreas with impaired glucose tolerance when compared with transgenic mice with intact ActR or littermates, but they showed the same alpha cell/beta cell ratios as their littermates. In addition to islet hypoplasia, the insulin response to glucose was severely impaired in dn-ActR transgenic mice. It is suggested that a precisely regulated intensity of activin signaling is necessary for the normal development of islets at the stage before differentiation into alpha and beta cells, and that activin plays a role in the postnatal functional maturation of islet beta cells.
Tomohiro Narita, Yasuo Ishii, Tsutomu Nohno, Sumihare Noji and Sadao Yasugi : Sonic hedgehog expression in developing chicken digestive organs is regulated by epithelial-mesenchymal interactions, Development Growth & Differentiation, Vol.40, No.1, 67-74, 1998.
(Summary)
Sonic hedgehog (Shh) gene encodes a secreted protein that acts as an important mediator of cell-cell interactions. A detailed analysis of Shh expression in the digestive organs of the chicken embryo was carried out. Shh expression in the endoderm begins at stage 7, when the formation of the foregut commences, and is found as narrow bands in the midgut. Shh expression around the anterior intestinal portal at stage 15 is restricted to the columnar endoderm lined by the thick splanchnic mesoderm, suggesting that the existence of thick splanchnic mesoderm might be necessary for Shh expression in the columnar endoderm. After the gut is closed, Shh expression is found universally in digestive epithelia, including the cecal epithelium. However, its expression ceases in the epithelium of the proventricular glands, the ductus choledochus and ductus pancreaticus that protrude from the main digestive duct. When the gizzard epithelium differentiated into glands under the influence of the proventricular mesenchyme, the glandular epithelium lost the ability to express Shh. These findings suggest that Shh expression in the epithelium may be regulated by surrounding mesenchyme throughout organogenesis of the digestive organs and is closely involved in epithelial-mesenchymal interactions in developing digestive organs.
Motomi Enomoto-Iwamoto, Masahiro Iwamoto, Yoshiki Mukudai, Yasuhiko Kawakami, Tsutomu Nohno, Yoshinobu Higuchi, Seiji Takemoto, Hideyo Ohuchi, Sumihare Noji and Kojiro Kurisu : Bone morphogenetic protein signaling is required for maintenance of differentiated phenotype, control of proliferation, and hypertrophy in chondrocytes, The Journal of Cell Biology, Vol.140, No.2, 409-418, 1998.
(Summary)
To examine the role of bone morphogenetic protein (BMP) signaling in chondrocytes during endochondral ossification, the dominant negative (DN) forms of BMP receptors were introduced into immature and mature chondrocytes isolated from lower and upper portions of chick embryo sternum, respectively. We found that control sternal chondrocyte populations expressed type IA, IB, and II BMP receptors as well as BMP-4 and -7. Expression of a DN-type II BMP receptor (termed DN-BMPR-II) in immature lower sternal (LS) chondrocytes led to a loss of differentiated functions; compared with control cells, the DN-BMPR- II-expressing LS chondrocytes proliferated more rapidly, acquired a fibroblastic morphology, showed little expression of type II collagen and aggrecan genes, and upregulated type I collagen gene expression. Expression of DN-BMPR-II in mature hypertrophic upper sternal (US) chondrocytes caused similar effects. In addition, the DN-BMPR-II-expressing US cells exhibited little alkaline phosphatase activity and type X collagen gene expression, while the control US cells produced both alkaline phosphatase and type X collagen. Both DN-BMPR-II-expressing US and LS chondrocytes failed to respond to treatment with BMP-2 . When we examined the effects of DN forms of types IA and IB BMP receptors, we found that DN-BMPR-IA had little effect, while DN-BMPR-IB had similar but weaker effects compared with those of DN-BMPR-II. We conclude that BMP signaling, particularly that mediated by the type II BMP receptor, is required for maintenance of the differentiated phenotype, control of cell proliferation, and expression of hypertrophic phenotype.
(Keyword)
Animals / Bone Morphogenetic Protein Receptors, Type I / Bone Morphogenetic Protein Receptors, Type II / Bone Morphogenetic Proteins / Cell Differentiation / Cell Division / Chick Embryo / Chondrocytes / Collagen / Phenotype / Protein-Serine-Threonine Kinases / Proteoglycans / Receptors, Cell Surface / Receptors, Growth Factor / Signal Transduction
Hideyo Ohuchi, Jun Takeuchi, Hidefumi Yoshioka, Yoshiyasu Ishimaru, Keiko Ogura, Naoki Takahashi, Toshihiko Ogura and Sumihare Noji : Correlation of wing-leg identity in ectopic FGF-induced chimeric limbs with the differential expression of chick Tbx5 and Tbx4., Development, Vol.125, No.1, 51-60, 1998.
132.
Hideyo Ohuchi, J Takeuchi, H Yoshioka, Y Ishimaru, K Ogura, N Takahashi, T Ogura and Sumihare Noji : FGF signalling is involved in differential expressions of Tbx5 and Tbx4 in the limb bud for determination of vertebrate limb identity, Development, Vol.125, 51-60, 1998.
133.
K. Iida, H. Koseki, H. Kakinuma, N. Kato, Y. Mizutani-Koseki, Hideyo Ohuchi, Hidefumi Yoshioka, Sumihare Noji, K. Kawamura, Y. Kataoka, F. Ueno, M. Taniguchi, N. Yoshida, T. Sugiyama and N. Miura : Essential roles of the winged helix transcription factor MFH-1 in aortic arch patterning and skeletogenesis, Development, Vol.124, No.22, 4627-4638, 1997.
(Summary)
Mesenchyme Fork Head-1 (MFH-1) is a forkhead (also called winged helix) transcription factor defined by a common 100-amino acid DNA-binding domain. MFH-1 is expressed in non-notochordal mesoderm in the prospective trunk region and in cephalic neural-crest and cephalic mesoderm-derived mesenchymal cells in the prechordal region of early embryos. Subsequently, strong expression is localized in developing cartilaginous tissues, kidney and dorsal aortas. To investigate the developmental roles of MFH-1 during embryogenesis, mice lacking the MFH-1 locus were generated by targeted mutagenesis. MFH-1-deficient mice died embryonically and perinatally, and exhibited interrupted aortic arch and skeletal defects in the neurocranium and the vertebral column. Interruption of the aortic arch seen in the mutant mice was the same as in human congenital anomalies. These results suggest that MFH-1 has indispensable roles during the extensive remodeling of the aortic arch in neural-crest-derived cells and in skeletogenesis in cells derived from the neural crest and the mesoderm.
Tsutomu Nohno, Yasuhiko Kawakami, Naoyuki Wada, Tetsuya Ishikawa, Hideyo Ohuchi and Sumihare Noji : Differential expression of the two closely related LIM-class homeobox genes LH-2A and LH-2B during limb development, Biochemical and Biophysical Research Communications, Vol.238, No.2, 506-511, 1997.
135.
N. Osumi, A. Hirota, Hideyo Ohuchi, M. Nakafuku, T. Iimura, S. Kuratani, M. Fujiwara, Sumihare Noji and K. Eto : Pax-6 is involved in the specification of hindbrain motor neuron subtype, Development, Vol.124, No.15, 2961-2972, 1997.
136.
Takashi Nakamura, Tomonao Aikawa, Motomi Iwamoto-Enomoto, Masahiro Iwamoto, Yoshinobu Higuchi, Pacifici Maurizio, Naoki Kinto, Akira Yamaguchi, Sumihare Noji, Kojiro Kurisu and Tokuzo Matsuya : Induction of osteogenic differentiation by hedgehog proteins, Biochemical and Biophysical Research Communications, Vol.237, No.2, 465-469, 1997.
137.
Hideyo Ohuchi, Takashi Nakagawa, Atsuyo Yamamoto, Akihiro Araga, Takashi Ohata, Yoshiyasu Ishimaru, Hidefumi Yoshioka, Takashi Kuwana, Tsutomu Nohno, M. Yamasaki, N. Itoh and Sumihare Noji : The mesenchymal factor, FGF10, initiates and maintains the outgrowth of the chick limb bud through interaction with FGF8, an apical ectodermal factor, Development, Vol.124, No.11, 2235-2244, 1997.
Mikiko Tanaka, Koji Tamura, Sumihare Noji, Tsutomu Nohno and Hiroyuki Ide : Induction of additional limb at the dorsal-ventral boundary of a chick embryo, Developmental Biology, Vol.182, No.2, 191-203, 1997.
Hideyo Ohuchi, Mami Shibusawa, Takashi Nakagawa, Takashi Ohata, Hidefumi Yoshioka, Yasokazu Hirai, Tsutomu Nohno, Sumihare Noji and Norio Kondo : A chick wingless mutation causes abnormality in maintenance of Fgf8 expression in the wing apical ridge, resulting in loss of the dorsoventral boundary, Mechanisms of Development, Vol.62, 3-13, 1997.
Nao Niwa, Mariko Saito, Hideyo Ohuchi, Hidefumi Yoshioka and Sumihare Noji : Corelation between Distal-less expression patterns and structures of of appendages in development of the two-spotted cricket, Gryllus bimaculatus, Zoological Science, Vol.14, No.1, 115-125, 1997.
E Koyama, L J Leatherman, Sumihare Noji and M Pacifici : Early chick limb cartilaginous elements possess polarizing acrivity and express Hedgehog-Related morphogenetic factors, Developmental Dynamics, Vol.207, 344-354, 1996.
Sachiko Iseki, Akihiro Araga, Hideyo Ohuchi, Tsutomu Nohno, Hidefumi Yoshioka, Yoshio Hayashi and Sumihare Noji : Sonic hedgehog is expressed in epithelial cells during development of whiskar, hair, and tooth, Biochemical and Biophysical Research Communications, Vol.218, No.3, 688-693, 1996.
Hidefumi Yoshioka, Hideyo Ohuchi, Y Ishimaru, T Ishikawa, Tsutomu Nohno, K Saigo and Sumihare Noji : A Drosophila receptor-type tyrosine kinase (DFR1) acts as a fibroblast growth factor receptor in Xenopus embryos, Development Growth & Differentiation, Vol.38, 617-624, 1996.
146.
Eiki Koyama, Tomoichiro Yamaai, Sachiko Iseki, Hideyo Ohuchi, Tsutomu Nohno, Hidefumi Yoshioka, Yoshio Hayashi, Judith L. Leatherman, Eleanor B. Golden, Sumihare Noji and Maurizio Pacifici : Polarizing activity, Sonic headgehog, and tooth development in embryonic and postnatal mouse, Developmental Dynamics, Vol.206, 59-72, 1996.
Tooth development / Polarizing activity / Sonic hedgehog / Enamel knot
147.
Tsutomu Nohno, Yasuhiko Kawakami, Hideyo Ohuchi, Akira Fujiwara, Hidefumi Yoshioka and Sumihare Noji : Involvement of the sonic hedgehog gene in chick feather formation, Biochemical and Biophysical Research Communications, Vol.206, No.1, 33-39, 1995.
Kohtaro Kawashima, Akira Yamaguchi, Toshimasa Shinki, Sumihare Noji, Satoshi Yokose, Tomoichiro Yamaai, Hiroyoshi Endo, Shusaku Yoshiki, Etsuko Abe and Tatsuo Suda : Microgravity generated by space flight has little effect on the growth and development of chick embryonic bone, Biol.Sci.Space, Vol.9, No.2, 82-94, 1995.
T Ishikawa, H Yoshioka, Hideyo Ohuchi, Sumihare Noji and T Nohno : Truncated type receptor for BMP-4 induces secondary axial structures in Xenopus embryos, Biochemical and Biophysical Research Communications, Vol.216, 26-33, 1995.
N Tanda, Y Kawakami, T Saito, Sumihare Noji and Tsutomu Nohno : Cloning and characterization of Wnt-4 and Wnt-11 cDNAs fro chick embryo, DNA Sequence, Vol.5, No.5, 277-281, 1995.
Tatsuo Mima, Hideyo Ohuchi, Sumihare Noji and Takashi Mikawa : FGF can induce outgrowth of somatic mesoderm both inside and outside of limb-forming regions, Developmental Biology, Vol.167, No.2, 617-620, 1995.
Fumio Myokai, N Washio, Y Asahara, Tomoichiro Yamaai, N Tanda, T Ishikawa, S Aoki, H Kurihara, Y Yamamura, T Saito, K Matsuo, T Nakamura, Sumihare Noji and Tsutomu Nohno : Expression of the hepatocyte growth factor gene during chick limb development, Developmental Dynamics, Vol.202, 80-90, 1995.
Sachiko Matsuhashi, Sumihare Noji, Eiki Koyama, Fumio Myokai, Hideyo Ohuchi, Shigehiko Taniguchi and Katsuji Hori : New gene,nel,encoding a Mr 93 K protein with EGF-like repeats is strongly expressed in neural tissues of earyl stage chick embryos, Developmental Dynamics, Vol.203, No.2, 212-222, 1995.
Hideyo Ohuchi, T Nakagawa, M Yamauchi, T Ohata, H Yoshioka, T Kuwana, T Mima, T Mikawa, T Nohno and Sumihare Noji : An additional limb can be induced from the flank of the chick embryo by FGF4, Biochemical and Biophysical Research Communications, Vol.209, 809-816, 1995.
Hideyo Ohuchi, H Yoshioka, A Tanaka, Y Kawakami, T Nohno and Sumihare Noji : Involvement of androgen-induced growth factor (FGF-8) gene in mouse embryogenesis and morphogenesis, Biochemical and Biophysical Research Communications, Vol.204, 882-888, 1994.
H Yoshioka, Hideyo Ohuchi, T Nohno, A Fujiwara, N Tanda, Y Kawakami and Sumihare Noji : Regional expression of the Cwnt-4 gene in developing chick central nervous system in relationship to the diencephalic neuromere D2 and a dorsal domain of the spinal cord, Biochemical and Biophysical Research Communications, Vol.203, 1581-1588, 1994.
Hideyo Ohuchi, E Koyama, F Myokai, T Nohno, F Shiraga, T Matsuo, N Matsuo, S Taniguchi and Sumihare Noji : Expression patterns of two fibroblast growth factor receptor genes during chick eye development, Eye Res, Vol.58, 649-658, 1994.
T Matsuo, N Osumi-Yamashita, Sumihare Noji, Hideyo Ohuchi, E Koyama, F Myokai, N Matsuo, S Taniguchi, H Doi, S Iseki, Y Ninomiya, M Fujiwara, T Watabe and K Eto : A mutation in the Pax-6 gene in rat small eye is associated with impaired migration of midbrain crest cells, Nature Genetics, Vol.3, No.4, 299-304, 1993.
T Nohno, E Koyama, F Myokai, S Taniguchi, Hideyo Ohuchi, T Saito and Sumihare Noji : A chicken homeobox gene related to Drosophila paired is predominantly expressed in the developing limb, Developmental Biology, Vol.158, 254-264, 1993.
Sumihare Noji, E Koyama, F Myokai, T Nohno, Hideyo Ohuchi, K Nishikawa and S Taniguchi : Differential expression of three FGF receptor genes, FGFR1, FGFR2 and FGFR3, in chick bone formation, Dentistry in Japan, Vol.29, 36-40, 1992.
Hideyo Ohuchi, Sumihare Noji, E Koyama, F Myokai, K Nishikawa, N Nohno, K Tashiro, K Shiokawa, N Matsuo and S Taniguchi : Expression pattern of the activin receptor type IIA gene during differentiation of chick neural tissues, muscle and skin., FEBS Letters, Vol.303, 185-189, 1992.
Taro Mito, Yoshiyasu Ishimaru, Takahito Watanabe, Taro Nakamura, Guillem Ylla, Sumihare Noji and G Cassandra Extavour : Cricket: The third domesticated insect., Current Topics in Developmental Biology, Vol.147, 291-306, Mar. 2022.
(Summary)
Many researchers are using crickets to conduct research on various topics related to development and regeneration in addition to brain function, behavior, and biological clocks, using advanced functional and perturbational technologies such as genome editing. Recently, crickets have also been attracting attention as a food source for the next generation of humans. In addition, crickets are increasingly being used as disease models and biological factories for pharmaceuticals. Cricket research has thus evolved over the last century from use primarily in highly important basic research, to use in a variety of applications and practical uses. These insects are now a state-of-the-art model animal that can be obtained and maintained in large quantities at low cost. We therefore suggest that crickets are useful as a third domesticated insect for scientific research, after honeybees and silkworms, contributing to the achievement of global sustainable development goals.
Tetsuya Bando, Taro Mito, Yoshimasa Hamada, Yoshiyasu Ishimaru, Sumihare Noji and Hideyo Ohuchi : Molecular mechanisms of limb regeneration: insights from regenerating legs of the cricket Gryllus bimaculatus, The International Journal of Developmental Biology, Vol.62, No.6-7-8, 559-569, Jun. 2018.
(Summary)
This review summarizes recent advances in leg regeneration research, focusing on the cricket Gryllus bimaculatus. Recent studies have revealed molecular mechanisms on blastema formation, establishment of positional information, and epigenetic regulation during leg regeneration. Especially, these studies have provided molecular bases in classical conceptual models such as the polar coordinate model, the intercalation model, the boundary model, the steepness model, etc., which were proposed to interpret regeneration processes of the cockroach legs. When a leg is amputated, a blastema is formed through the activation of the Janus-kinase (Jak)/Signal-Transduction-and-Activator-of-Transcription (STAT) pathway. Subsequently, the Hedgehog/Wingless/Decapentaplegic/Epidermal-growth-factor pathways instruct distalization in the blastema, designated as the molecular boundary model. Downstream targets of this pathway are transcription factors Distal-less (Dll) and dachshund (dac), functioning as key regulators of proximodistal pattern formation. Dll and dac specify the distal and proximal regions in the blastema, respectively, through the regulation of tarsal patterning genes. The expression of leg patterning genes during regeneration may be epigenetically controlled by histone H3K27 methylation via Enhancer-of-zeste and Ubiquitously-transcribed-tetratricopeptide-repeat-gene-X-chromosome. For the molecular mechanism of intercalation of the missing structures between the amputated position and the most distal one, Dachsous/Fat (Ds/Ft) steepness model has been proposed, in which the Ds/Ft pathway maintains positional information and determines leg size through dac expression. This model was theoretically verified to interpret the experimental results obtained with cricket legs. Availability of whole-genome sequence information, regeneration-dependent RNA interference, and genome editing technique will have the cricket be an ideal model system to reveal gene functions in leg regeneration.
中尾 一和 and Sumihare Noji : 再生医学, The Medical Frontline, 51-59, Dec. 2000.
13.
Sumihare Noji and Hideyo Ohuchi : Functions of fibroblast growth factor and their receptors during morphogenesis, Tanpakushitsu Kakusan Koso, Vol.45, No.13, 2094-2099, Sep. 2000.
14.
Sumihare Noji : 形態形成遺伝子の臨床利用をめざして, New Horizon for Medicine, Vol.32, No.8, 22-28, Aug. 2000.
15.
直人 上野, 岡野 栄之 and Sumihare Noji : 発生·神経研究の最前線2000, Experimental Medicine, Vol.18, No.9, 64-68, May 2000.
Hideyo Ohuchi and Sumihare Noji : FGF-induced additional limbs in the study of initiation of limb formation, limb identity, myogenesis, and innervation, Cell and Tissue Research, Vol.296, 45-56, 1999.
T Nohno, Hideyo Ohuchi, H Yoshioka and Sumihare Noji : Involvement of growth factors and their receptors in morphogenesis of the chick limb, Gann Monograph on Cancer Research, Vol.42, 79-90, 1994.
29.
Hideyo Ohuchi and Sumihare Noji : 発生分化におけるレチノイン酸の役割, The Cell, Vol.25, 423-427, 1993.
Shintaro Inoue, Takahito Watanabe, Hamaguchi Taiki, Fujie Kai, Shimamura Ayane, Yoshiyasu Ishimaru, Katsuyuki Miyawaki, Takeshi Nikawa, Akira Takahashi, Sumihare Noji and Taro Mito : Artificial modification of cricket body color: breeding for the next-generation of protein supply, International Conference of Non-Traditional Arthropod Model Systems, Aug. 2023.
2.
Fujie Kai, Shintaro Inoue, Hamaguchi Taiki, Yoshiyasu Ishimaru, Katsuyuki Miyawaki, Takeshi Nikawa, Akira Takahashi, Sumihare Noji, Takahito Watanabe and Taro Mito : The discovery of two paralogous dopamine-synthase genes in the two-spotted cricket Gryllus bimaculatus, International Conference of Non-Traditional Arthropod Model Systems, Aug. 2023.
3.
Silvia Naomi Mitsui Akagi, Akihiro Yasue, Kiyoshi Masuda, Takuya Naruto, Seiichi Oyadomari, Sumihare Noji, Issei Imoto and Eiji Tanaka : Conserved C-terminal domain of MSX1 is essential for tooth development, 12th Tooth Morphogenesis and Differentiation conference, Porvoo, Finland, Jun. 2016.
4.
Matsuoka Yuji, Takahito Watanabe, Sayuri Tomonari, Sumihare Noji and Taro Mito : Functional analysis of a Hox gene, abdominal-A, using CRISPR/Cas9 system in the cricket Gryllus bimaculatus, International Tribolium Meeting 2015, Berkeley, USA, Aug. 2015.
5.
Takahito Watanabe, Matsuoka Yuji, Sayuri Tomonari, Kurita Chinami, Sumihare Noji and Taro Mito : Genome editing in the two-spotted cricket, Gryllus bimaculatus, using CRISPR/Cas9 system, Insect Genetic Technologies Workshop, Manhattan, Kansas, USA, Jun. 2015.
6.
Taro Mito, Itoh Takehiko, Morimoto Hiroya, Kajitani Ray, Toyoda Atsushi, Sayuri Tomonari, Fuketa Masao, Takahito Watanabe, Matsuoka Yuji and Sumihare Noji : Genome sequencing and annotation of the cricket Gryllus bimaculatus, a hemimetabolous insect model, Ninth Annual Arthropod Genomics Symposium, Manhattan, Kansas, USA, Jun. 2015.
7.
Akihiro Yasue, Silvia Naomi Mitsui Akagi, Teppei Watanabe, T Sakuma, Seiichi Oyadomari, T Yamamoto, Sumihare Noji, Taro Mito and Eiji Tanaka : Highly efficient targeted mutagenesis in one-cell mouse embryos mediated by TALEN and CRISPR/Cas systems, X meeting for Spanish Society for Developmental Biology (SEBD), Madrid, Oct. 2014.
8.
Sumihare Noji, Taro Mito, Bando Tetsuya, Nakamura Taro, Takahito Watanabe, Ishimaru Yoshiyasu and Hideyo Ohuchi : Regeneration of insect legs from stem cells, Thirteenth International Congress on Invertebrate Reproduction and Development, Detroit, Detroit, MI, USA, Jul. 2014.
9.
Nakamura Taro, Sumihare Noji and Taro Mito : Molecular mechanisms underlying early embryonic patterning and germ cell specification in the cricket, International Symposium on RNAi and Genome editing methods, Tokushima, Japan, Mar. 2014.
10.
Takahito Watanabe, Sumihare Noji and Taro Mito : Targeted genome modifications in the cricket, Gryllus bimaculatus, using CRISPR/Cas9 system, International Symposium on RNAi and Genome editing methods, Tokushima, Japan, Mar. 2014.
11.
Akihiro Yasue, Silvia Naomi Mitsui Akagi, H Watanabe, T Sakuma, Seiichi Oyadomari, T Yamamoto, Sumihare Noji, Taro Mito and Eiji Tanaka : A high efficient gene targeting in one-cell mouse embryos mediated by TALEN and CRISPR/Cas system., International Symposium on RNAi and Genome Editing Methods, Tokushima, Mar. 2014.
12.
Taro Mito, Takahito Watanabe and Sumihare Noji : Genome modification technology in the cricket Gryllus bimaculatus, 1st Asian Invertebrate Immunity Symposium, Busan, Feb. 2014.
13.
Takahito Watanabe, Matsuoka Yuji, Sumihare Noji and Taro Mito : Targeted genome editing in the cricket, Gryllus bimaculatus, using CRISPR/Cas9 system, FASEB SRC on Genome Engineering-Cutting-Edge Research and Applications, Nassau, Bahamas, Jan. 2014.
14.
Takahito Watanabe, Ochiai Hiroshi, Sakuma Tetsushi, Ishihara Satoshi, Nakamura Taro, Yamamoto Takashi, Sumihare Noji and Taro Mito : Targeted genome modifications in the cricket, Gryllus bimaculatus, Conference of Transposition & Genome Engineering 2013, Budapest, Hungary, Sep. 2013.
15.
Takahito Watanabe, Hiroshi Ochiai, Tetsushi Sakuma, Hadley W. Horch, Naoya Hamaguchi, Taro Nakamura, Tetsuya Bando, Hideyo Ohuchi, Takashi Yamamoto, Sumihare Noji and Taro Mito : Gene knockout in a hemimetabolous insect Gryllus bimaculatus by nontransgenic genome modification with zinc-finger and TALE nucleases, Asia-Pacific Developmental Biology Conference, Taipei, Taiwan, Oct. 2012.
16.
Takahito Watanabe, Hiroshi Ochiai, Tetsushi Sakuma, Taro Nakamura, Taro Mito, Takashi Yamamoto and Sumihare Noji : Efficient production of knockout crickets using custom designed nucleases, ZFNs and TALENs, FASEB Science Research Conferences: Genome Engineering; Research & Applications, Lucca, Italy, Sep. 2012.
17.
Taro Mito, Nakamura Taro, Bando Tetsuya, Watanabe Takahito and Sumihare Noji : Exploring mechanisms of embryonic patterning in Gryllus bimaculatus, a hemimetablous insect model system, [Symposium: From embryo to metamorphosis: Genes for insect development (Organizers: Sumihare Noji and Martin Klingler)], 24th International Congress of Entomology, Daegu, Korea, Aug. 2012.
18.
Tetsuya Bando, Yoshimasa Hamada, Taro Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Dachsous/Fat signaling via Hippo/Salvador/Warts pathway regulates cell proliferation and pattern formation during leg regeneration in the cricket, 24th International Congress of Entomology, Daegu, Korea, Aug. 2012.
19.
Taro Nakamura, Taro Mito, Tetsuya Bando and Sumihare Noji : Role of Wnt and BMP signaling pathways in the regional specification of early blastoderm in the cricket Gryllus bimaculatus,, 24th International Congress of Entomology, Daegu, Korea, Aug. 2012.
20.
T Watanabe, H Ochiai, T Sakuma, T Nakamura, Taro Mito, Hideyo Ohuchi, T Yamamoto and Sumihare Noji : Efficient production of knockout crickets using zinc-finger nucleases, The 2nd International Conference on the Cricket / RNAi Symposium, tokushima,Japan, Mar. 2012.
21.
T Nakamura, Taro Mito, T Bando and Sumihare Noji : Involvement of Wnt and BMP signaling pathways in the regional specification of early blastoderm in the cricket Gryllus bimaculatus, The 2nd International Conference on the Cricket / RNAi Symposium, tokushima,Japan, Mar. 2012.
22.
T Bando, Y Matsuoka, Y Hamada, T Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Molecular mechanisms underlying cell proliferation and pattern formation during leg regeneration in Gryllus bimaculatus, The 2nd International Conference on the Cricket / RNAi Symposium, tokushima,Japan, Mar. 2012.
23.
Y Matsuoka, T Bando, T Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Functional analysis of epigenetic regulation during embryogenesis of the cricket, Gryllus bimaculatus, The 2nd International Conference on the Cricket / RNAi Symposium, tokushima,Japan, Mar. 2012.
24.
Makoto Mizunami, T Takahashi, A Hamada, Katsuyuki Miyawaki, Y Matsumoto, Taro Mito and Sumihare Noji : Systemic RNA interference for the study of long-term memory formation in the cricket, The 2nd International Conference on the Cricket / RNAi Symposium, tokushima,Japan, Mar. 2012.
25.
Taro Mito, T Nakamura, T Watanabe, H Ochiai, T Sakuma, T Bando, Hideyo Ohuchi and Sumihare Noji : Exploring mechanisms of embryonic patterning in Gryllus bimaculatus, a hemimetabolous insect model system, The 2nd International Conference on the Cricket / RNAi Symposium, tokushima,Japan, Mar. 2012.
26.
Taro Mito, T Nakamura, T Watanabe, T Band and Sumihare Noji : Functional genomics of the cricket Gryllus bimaculatus, a model system for regeneration and evolutionary developmental studies, 5th Annual Arthropod Genomics Symposium, Kansas City,USA, Jun. 2011.
27.
Taro Mito, T Nakamura, T Bando and Sumihare Noji : Ancestral developmental mechanisms in insects revealed by RNAi analysis of cricket genes, [Symposium: RNA interference- comparative studies of gene functions in invertebrates, 8th International Congress on Comparative Physiology and Biochemistry, Nagoya, Jun. 2011.
28.
Hideyo Ohuchi, Mayumi Okamoto, Akane Matsuyo, Takumi Kawaue, Sayuri Tomonari and Sumihare Noji : Generation of neural retina from pigmented retinal progenitors by an intrinsic factor Lhx1, Sesimbra, Portugal, Sep. 2010.
Emi Kawakami, Nao Kinouchi, Adachi Taro, Yutaka Ohsawa, Naozumi Ishimaru, Hideyo Ohuchi, Yoshihide Sunada, Yoshio Hayashi, Eiji Tanaka and Sumihare Noji : Special Processed Collagen-mediated Application of Myostatin-siRNA for Muscular Atrophy Diseases, 88th IADR, Barcelona, Jul. 2010.
31.
Hideyo Ohuchi and Sumihare Noji : Regulation of leg size and shape by dachshund, Distal-less, and the Dachsous/Fat signaling pathway during cricket leg regeneration, the College of William and Mary in Williamsburg, Virginia. USA, Jul. 2010.
32.
T Nakamura, Taro Mito, M Yoshizaki, T Bando and Sumihare Noji : Dynamic control of positional specification in a primitive mode of insect segmentation, フランス(パリ第7大学), Jul. 2010.
33.
Sumihare Noji, Tetsuya Bando, Taro Mito, Taro Nakamura, Takahito Watanabe and Hideyo Ohuchi : Regulation of leg size and shape by the Dachsous/Fat signalling pathway during cricket leg regeneration, Washington, DC, USA, Apr. 2010.
34.
Taro Mito, Taro Nakamura, Masato Yoshizaki, Tetsuya Bando, Hideyo Ohuchi and Sumihare Noji : Highly dynamic cell behavior during early development in the intermediate germ insect Gryllus bimaculatus, as revealed by analyses of transgenic embryos, Washington DC, USA, Apr. 2010.
35.
Emi Kawakami, Nao Kinouchi, Yutaka Osawa, Naozumi Ishimaru, Hideyo Ohuchi, Yoshihide Sunada, Yoshio Hayashi, Eiji Tanaka and Sumihare Noji : Strategic study of atelocollagen-mediated application of mystatin-targeting siRNA for therapeutic use for muscular atrophy diseases, QOL International Congress, Niigata, Feb. 2010.
H Tao, M Shimizu, R Kusumoto, K Ono and Sumihare Noji : Roles of FGF10 in mouse eyelid development., 15th International Society of Developmental Biologists Congress, Sydney, Australia, Sep. 2005.
(Keyword)
FGF10
38.
Hideyo Ohuchi, T Adachi, Y Hayashibara, M Mitsumori, T Ohata, Y Kawano, M Ogasawara and Sumihare Noji : Novel in situ hybridization methods with InSitu Chip and Sheet-Chip to construct expression database for all mouse genes, 10th Anniversary of Kazusa DNA Research Institute 14th International Workshop, Japan, Oct. 2004.
Taro Mito, Katsuyuki Miyawaki, Y Shinmyo, Hideyo Ohuchi and Sumihare Noji : Genome-wide RNAi in the cricket Gryllus bimaculactus as a new model system to study gene functions, 10th Anniversary of Kazusa DNA Research Institute 14th International Workshop, Japan, Oct. 2004.
Sumihare Noji, Taro Mito, Y Tanaka, Katsuyuki Miyawaki, (名) Shinmyo and Hideyo Ohuchi : Involvement of Wnt and EGF signaling systems in regeneraion of cricket legs, 8th international Conference Limb Development and Regeneration, 47, Dundee, UK, Jul. 2004.
41.
Hideyo Ohuchi, H Kurose, H Tao and Sumihare Noji : Gene expression in the developing chicken retina by EST sequencing and in situ hybridization analysis, Gordon Research Conference on Visual System Development, USA, Jun. 2002.
42.
F Saito, T Manabe, Y Miyatake, N Itoh, Sumihare Noji and Hideyo Ohuchi : Roles of mesenchymal FGFs in chick limb development, 14th International Congress of Developmental Biology, Japan, Jul. 2001.
43.
H Sasaki, T Yamaoka, A Yasue, Hideyo Ohuchi, M Itakura, S Kato, M Nagayama and Sumihare Noji : Expression pattern of Fgf10 and its regulatory element in oral tissues, 14th International Congress of Developmental Biology, Japan, Jul. 2001.
44.
A Yasue, H Tao, T Nohno, K Moriyama, Sumihare Noji and Hideyo Ohuchi : Cloning and expression of the chick p63 gene, 14th International Congress of Developmental Biology, Japan, Jul. 2001.
45.
H Tao, H Yoshioka, T Nohno, Sumihare Noji and Hideyo Ohuchi : Roles for FGF10 as a mesenchymal factor during chick feather bud formation, 14th International Congress of Developmental Biology, Japan, Jul. 2001.
46.
Sumihare Noji : Involvement of developmental regulatory genes in leg regeneration in a hemimetabolous insect (cricket), 14th international congress of developmental biology, Kyoto, Japan, Jul. 2001.
47.
Sumihare Noji : Limb development of the hemimetabolous short-germ insect, Gryllus bimaculatus (cricket), ENBO Workshop:Embryonic organizer signaling: the next frontiers, Heidelberg, Germany, Apr. 2001.
48.
Sumihare Noji : Comparative study on development and regeneration of the cricket leg, International symposium in Okazaki: new horizons of developmental biology, Okazaki, Japan, Mar. 2001.
49.
Sumihare Noji : FGF10 is involved in budding and branching morphogenesis as a mesenchymal factor in epithelial-mesenchymal interaction, Second NHRI conference on developmental biology, Taoyuan, Taiwan, Oct. 2000.
50.
Sumihare Noji, Yoshiko Inoue, Taro Mito and Hideyo Ohuchi : Limb development and regeneration of the cricket leg: Expression patterns of hedgehog, wingless and decepentaplegic, 7th International conference on limb development and regeneration, Vol.90, No.1, 20, Aussois, France, Aug. 2000.
51.
Sumihare Noji, Y Inoue, Taro Mito and Hideyo Ohuchi : Limb development and Regeneration of the cricket leg: Expression patterns of hedgehog, wingless and decapentaplegic, 7th International Conference on Limb Development and Regeneration, France, May 2000.
52.
Hideyo Ohuchi, F Saitoh, N Itoh and Sumihare Noji : Roles of FGF18 in chick limb development, 7th International Conference on Limb Development and Regeneration, France, May 2000.
53.
Sumihare Noji : Molecular mechanism of pattern formation of the limb, The 5th international symposium on congenital differences of the upper limb, 5, Kyoto, May 2000.
54.
Sumihare Noji : FGF10, a mesenchymal factor involved in budding morphogenesis through epithelial-mesenchymal interaction, EMBO Workshop: FGFs and their receptors: structure to function, Ein Gedi, Dead Sea, Israel, Dec. 1999.
55.
Hideyo Ohuchi, S Tomonari, H Itoh, T Mikawa and Sumihare Noji : Identification of chick rax/rx genes with overlapping patterns of expression during early eye and brain development, International meeting of ISDB (International Society of Developemtal Biologists), Norway, Jun. 1999.
56.
Sumihare Noji : Molecular mechanisms of vertebrate limb initiation and specification, The 4th conference of the international federation of placental associations, Tokyo, Oct. 1998.
57.
Hideyo Ohuchi, Sumihare Noji and N Itoh : Novel fibroblast growth factors in developing nervous systems, The 12th Biennial Meeting of International Society for Developmental Neurobiology (ISDN), Canada, Aug. 1998.
58.
Sumihare Noji : Molecular mechanisms for vertebrate limb initiation and specification, French-Japanese worksop: Genes and early development, Lyon, France, Jun. 1998.
59.
Hideyo Ohuchi, H Yoshioka, M Yamasaki, H Itoh, N Itoh, T Nohno and Sumihare Noji : Molecular mechanisms for vertebrate limb initiation and specification, A Keystone Symposium Vertebrate Development, USA, Apr. 1998.
60.
Hideyo Ohuchi, H Yoshioka, M Yamasaki, N Itoh, T Nohno and Sumihare Noji : A mesenchymal factor of FGF10 initiates and maintains the outgrowth of the chick limb bud through interactions with an apical ectodermal factor of FGF8, The 13th International Congress of Developmental Biology, USA, Jul. 1997.
61.
Hideyo Ohuchi, T Nakagawa, T Ohata, H Tanaka, T Nohno, H Yoshioka and Sumihare Noji : Molecular analysis of the extra limb ("Dasoku") induced by FGF4 in the chick embryonic flank, The 12th International Symposium of Federation of Asian Oceanian Biochemists and Molecular Biologists (FAOBMB), Japan, Jul. 1996.
62.
Hideyo Ohuchi, H Tanaka, T Nohno, T Ishikawa, Y Kawakami, H Yoshioka and Sumihare Noji : Molecular analysis of the extra limb ("Dasoku") induced by FGF4 in the chick embryonic flank, The 5th International Limb Development and Regeneration Conference, U.K, Mar. 1996.
63.
Hideyo Ohuchi, H Tanaka, T Kuwana, T Mikawa, T Nohno, H Yoshioka and Sumihare Noji : Induction of an additional limb by retroviral ectopic expression of the Fgf4 gene in chicken embryos, The 37th NIBB (National Institute for Basic Biology) Conference, Okazaki,Japan, Mar. 1996.
64.
Hideyo Ohuchi, T Nakagawa, M Yamauchi, T Ohata, T Kuwana, T Mima, T Mikawa, T Nohno, H Yoshioka and Sumihare Noji : An additional limb can be induced from the flank of the chick embryo by FGF4, The 3rd International Duodecim Symposium, Finland, Jun. 1995.
65.
Hideyo Ohuchi, T Mima, H Tanaka, H Yoshioka, T Nohno, Sumihare Noji and T Mikawa : Roles of the FGF and Wnt families in chick development, British Society for Developmental Biology, U.K, Apr. 1994.
Higashihara Aya, Yoshiyasu Ishimaru, Matsumura Saki, Kawamoto Kohei, Sayuri Tomonari, Sumihare Noji and Taro Mito : Gene knock-out analysis of a metamorphosis factor Myoglianin in the cricket Gryllus bimaculatus, The 43rd Annual Meeting of the Molecular Biology Society of Japan, Online, Dec. 2020.
Takuya Watari, Yoshiyasu Ishimaru, Sumihare Noji and Taro Mito : Involvement of macrophages in leg regeneration of the cricket Gryllus bimaculatus, 52nd Annual Meeting of the Japanese Society of Developmental Biologists, May 2019.
9.
Kohei Kawamoto, Mayuko Matsuda, Takahisa Yamashita, Takahito Watanabe, Sayuri Tomonari, Yoshiyasu Ishimaru, Sumihare Noji and Taro Mito : Precise in-frame integration of a GFP gene using microhomology-mediated knock-in technology in Gryllus bimaculatus, 52nd Annual Meeting of the Japanese Society of Developmental Biologists, May 2019.
10.
Yuki Nakamura, Sayuri Tomonari, Kohei Kawamoto, Takahito Watanabe, Yoshiyasu Ishimaru, Sumihare Noji and Taro Mito : Resolving the correlation between phenotype and genotype in a segmentation gene even-skipped in the cricket Gryllus bimaculatus, 52nd Annual Meeting of the Japanese Society of Developmental Biologists, May 2019.
11.
Yu-ki Nakamura, Ko-hei Kawamoto, Sayuri Tomonari, Takahito Watanabe, Yoshiyasu Ishimaru, Taro Mito and Sumihare Noji : Gene knock-out analysis of a segmentation gene even-skipped in the cricket Gryllus bimaculatus, Joint Annual Meeting of JSDB 51st and JSCB 70th, Jun. 2018.
Matsuda Mayuko, Matsuoka Yuji, Yoshiyasu Ishimaru, Sayuri Tomonari, Takahito Watanabe, Sumihare Noji and Taro Mito : Functional analysis of a Hox gene, abdominal-A, in the cricket Gryllus bimaculatus using a CRISPR/Cas9-mediated gene knock-in system, Japanese Society of Developmental Biologists, May 2017.
14.
Nakamura Yu-Ki, Kawamoto Kohei, Sayuri Tomonari, Matsuda Mayuko, Takahito Watanabe, Yoshiyasu Ishimaru, Uemura Natsuki, Sumihare Noji and Taro Mito : even-skipped is required for segmentation and elongation of embryos in the cricket Gryllus bimaculatusas revealed by CRISPR/Cas9-based gene knock-out., Japanese Society of Developmental Biologists, May 2017.
15.
Kohei Kawamoto, Sayuri Tomonari, Yuji Matsuoka, Takahito Watanabe, Yoshiyasu Ishimaru, Sumihare Noji and Taro Mito : even-skipped acts principally as a gap gene in the cricket Gryllus bimaculatus as revealed by CRISPR/Cas9-based gene knockout analysis, JSDB Special Symposium: Frontier of Developmental Biology Hosted by JSDB, Jun. 2016.
Silvia Mitsui, Akihiro Yasue, Issei Imoto, Seiichi Oyadomari, Sumihare Noji, Taro Mito and Eiji Tanaka : In vivo study of Msx1 gene in mice using CRISPR/Cas system, 第47回日本発生生物学会, May 2014.
25.
Yuji Matsuoka, Tetsuya Bando, Takahito Watanabe, Sumihare Noji and Taro Mito : Functions of Polycomb group gene in regulation of Hox gene expression in a primitive mode of insect embryogenesis in the cricket Gryllus bimaculatus, 第47回日本発生生物学会, May 2014.
26.
Yoshimasa Hamada, Tetsuya Bando, Taro Mito, Kenji Tomioka, Sumihare Noji and Hideyo Ohuchi : Epigenetic regulation of gene expressions via methylation on histone H3 27th lysine residue during leg regeneration, 第47回日本発生生物学会, May 2014.
27.
Takahito Watanabe, Yuji Matsuoka, Taro Mito and Sumihare Noji : Targeted gene disruption in the cricket, Gryllus bimaculatus, using CRISPR/Cas9 system, 第47回日本発生生物学会, May 2014.
28.
Taro Mito, 渡辺 崇人 and Sumihare Noji : CRISPR/Casシステムを用いたフタホシコオロギにおけるゲノム編集, 第58回日本応用動物昆虫学会大会, Mar. 2014.
Akihiro Yasue, Silvia Naomi Mitsui Akagi, Teppei Watanabe, T Sakuma, Seiichi Oyadomari, T Yamamoto, Sumihare Noji, Taro Mito and Eiji Tanaka : A high efficient gene targeting in one-cell mouse embryos mediated by TALEN and CRISPR/Cas system, 第36回日本分子生物学会, Dec. 2013.
31.
Akihiro Yasue, Silvia Naomi Mitsui Akagi, Teppei Watanabe, T Sakuma, Seiichi Oyadomari, T Yamamoto, Sumihare Noji, Taro Mito and Eiji Tanaka : A high efficient gene targeting in one-cell mouse embryos mediated by TALEN and CRISPR/Cas system, 第36回日本分子生物学会, Dec. 2013.
渡辺 崇人, 松岡 佑児, 石原 聡, 山本 卓, Sumihare Noji and Taro Mito : ゲノム編集技術によるノックアウトコオロギの作製, 第15回日本進化学会大会, Aug. 2013.
36.
Nakamura Taro, Sumihare Noji and Taro Mito : Regulation of Wnt and BMP signaling pathways in the regional specification of early blastoderm in the cricket Gryllus bimaculatus, 第46回日本発生生物学会年会, May 2013.
37.
Bando Tetsuya, Taro Mito, Ohuchi Hideyo and Sumihare Noji : JAK/STAT signaling promotes blastemal cell proliferation during leg regeneration in the cricket Gryllus bimaculatus, 第46回日本発生生物学会年会, May 2013.
38.
松岡 佑児, 板東 哲哉, 中村 太郎, 渡辺 崇人, Sumihare Noji and Taro Mito : Epigenetic regulation of Hox gene expression by PcG genes in a primitive mode of insect embryogenesis in the cricket Gryllus bimaculatus, 第46回日本発生生物学会年会, May 2013.
39.
Takahito Watanabe, Hiroshi Ochiai, Tetsushi Sakuma, Taro Nakamura, Taro Mito, Takashi Yamamoto and Sumihare Noji : Targeted genome modifications using ZFNs and TALENs in the cricket Gryllus bimaculatus, 第46回日本発生生物学会年会, May 2013.
40.
T Bando, Y Ishimaru, T Kida, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Molecular mechanisim of regulation of blastemal cell proliferation during leg regeneration in Gryllus bimaculatus, 第35回日本分子生物学会年会, Dec. 2012.
41.
Takahito Watanabe, Hiroshi Ochiai, Tetsushi Sakuma, Taro Nakamura, Taro Mito, Takashi Yamamoto and Sumihare Noji : Generation of knockout crickets using ZFNs and TALENs, 第35回日本分子生物学会年会, Dec. 2012.
Takahito Watanabe, Hiroshi Ochiai, Tetsushi Sakuma, Taro Nakamura, Taro Mito, Hideyo Ohuchi, Takashi Yamamoto and Sumihare Noji : Efficient production of knockout crickets using zinc-finger nucleases, 第45回日本発生生物学会年会, May 2012.
44.
Tetsuya Bando, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Angiomotin regulates cell proliferation cooperatively with Expanded and Merlin during leg regeneration in Gryllus bimaculatus, 第45回日本発生生物学会年会, May 2012.
45.
Taro Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Regulation of orthodenticle and Wnt/Cad signaling pathway in anterior-posterior axis patterning during cricket early embryogenesis, 第45回日本発生生物学会年会, May 2012.
46.
Sumihare Noji, T Bando, Taro Mito and Hideyo Ohuchi : Molecular mechanisms underlying insect leg regeneration: from wound healing to leg size determination, 第34回日本分子生物学会年会, 2011.
47.
Y Matsuoka, T Bando, T Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Polycomb group genes epigenetically determines segmental identity in the cricket, Gryllus bimaculatus, 第34回日本分子生物学会年会, Dec. 2011.
48.
T Bando, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Angiomotin regulates leg size cooperatively with Expanded and Merlin during regeneration in Gryllus bimaculatus, 第34回日本分子生物学会年会, Dec. 2011.
49.
T Watanabe, H Ochiai, T Sakuma, T Nakamura, Taro Mito, Hideyo Ohuchi, T Yamamoto and Sumihare Noji : Making knockout crickets with zinc-finger nucleases, 第34回日本分子生物学会年会, Dec. 2011.
50.
Taro Mito, T Nakamura, T Watanabe, H Ochiai, T Sakuma, T Bando, Hideyo Ohuchi and Sumihare Noji : Exploring molecular mechanisms of early embryogenesis in the cricket Gryllus bimaculatus, 第34回日本分子生物学会年会, Dec. 2011.
51.
A Nakai, M Yoshizaki, Taro Mito, T Nakamura, T Bando, Hideyo Ohuchi and Sumihare Noji : Role of the orthodenticle gene in an ancestral mode of insect embryogenesis, as revealed by expression and functional analyses in the cricket Gryllus bimaculatus, 第44回日本発生生物学会年会, May 2011.
52.
T Nakamura, Taro Mito, M Yoshizaki, A Nakai, Hideyo Ohuchi and Sumihare Noji : Imaging of transgenic cricket embryos reveals cell movements consist with a syncytial patterning mechanism, 第44回日本発生生物学会年会, May 2011.
53.
Y Hamada, T Bando, Y Matsuoka, T Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Epigenetic regulation of gene expressions during leg regeneration in the two-spotted cricket, Gryllus bimaculatus, 第44回日本発生生物学会年会, May 2011.
54.
T Bando, Y Hamada, T Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Regulatory mechanism of blastemal cells mediated by polarity complexes via Dachsous/Fat and Hippo/Salvador/Warts pathway during leg regeneration in Gryllus bimaculatus, 第44回日本発生生物学会年会, May 2011.
55.
T Watanabe, H Ochiai, T Sakuma, M Asahina, T Nakamura, Taro Mito, Hideyo Ohuchi, T Yamamoto and Sumihare Noji : Targeted manipulation of genes with zinc finger nucleases in the cricket, Gryllus bimaculatus, 第44回日本発生生物学会年会, May 2011.
56.
K Matsuda, T Nakamura, T Bando, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Spatio-temporally controlled misexpression of genes using the GAL4/UAS system in the cricket, Gryllus bimaculatus, 第44回日本発生生物学会年会, May 2011.
Takahito Watanabe, H Ochiai, T Sakuma, Miki Asahina, Taro Nakamura, Taro Mito, Hideyo Ohuchi, T Yamamoto and Sumihare Noji : Targeted manipulation of genes with zinc finger nucleases in the cricket, Gryllus bimaculatus, 第33回日本分子生物学会年会, Dec. 2010.
59.
Kazuki Kurita, Akira Takagi, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Functions of the Drosophila retinal determination gene homologoues in eye development of a Hemimetabolous insect Gryllus bimaculatus, 第33回日本分子生物学会年会, Dec. 2010.
60.
Tetsuya Bando, Yuji Matsuoka, Yoshimasa Hamada, Taro Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Epigenetic regulation of gene expressions during leg regeneration in the two-spotted cricket Gryllus bimaculatus, 第33回日本分子生物学会年会, Dec. 2010.
61.
Masato Yoshizaki, Taro Nakamura, Taro Mito, Tetsuya Bando, Takahito Watanabe, Hideyo Ohuchi and Sumihare Noji : Functions of the orthodenticle-related genes during embryogenesis in the cricket Gryllus bimaculatus, 第33回日本分子生物学会年会, Dec. 2010.
Koji Matsuda, Taro Nakamura, F Ito, Taro Mito, Tetsuya Bando and Sumihare Noji : Development of enhancer trap lines using the GAL4/UAS system in the cricket, Gryllus bimaculatus, 第43回日本発生生物学会年会, Jun. 2010.
66.
Noha Dabour, Tetsuy Bando, Taro Nakamura, Katsuyuki Miyawaki, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Control of body size by chico and epidermal growth factor receptor, as revealed by systemic nymphal RNA interference in the cricket, 第43回日本発生生物学会年会, Jun. 2010.
67.
Yuji Matsuoka, Tetsuya Bando, Taro Nakamura, Hideyo Ohuchi, Sumihare Noji and Taro Mito : Enhancer of zeste epigenetically regulates leg development in the cricket Gryllus bimaculatus, 第43回日本発生生物学会年会, Jun. 2010.
68.
T Nakamura, Taro Mito, T Bando, Hideyo Ohuchi and Sumihare Noji : Dynamic control of positional specification in a primitive mode of insect segmentation, 第43回日本発生生物学会年会, Jun. 2010.
69.
Kazuki Kurita, Yohei Shinmyo, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Divergent function of Delta/Notch signaling in formation of body segments in the intermediate-germband cricket Gryllus bimaculatus, 第43回日本発生生物学会年会, Jun. 2010.
70.
Tetsuya Bando, Taro Nakamura, Taro Mito, Hideyo Ohuchi and Sumihare Noji : Lowfat regulates leg size and growth under the Dachsous/Fat signaling during regeneration in Gryllus bimaculatus, 43rd Annual Meeting for the Japanese Society of Developmental Biologists, Jun. 2010.
Analysis of Pancreatic B-Cell Development and an Animal Model of Regeneration Therapy for Diabetes Mellitus Using Transgenic Mice (Project/Area Number: 13671194 )
Mechanism of evolution and structure of developmental systems (Project/Area Number: 13044001 )
DEVELOPMENT OF A NEW THERAPEUTC APPROCH TO BONE REGENERATION FOR GENE THERAPY (Project/Area Number: 12557153 )