{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/40790037","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467276","label":"url"}],"paper_title":{"en":"Unique metabolic regulation of micromeres contributes to gastrulation in the sea urchin embryo.","ja":"Unique metabolic regulation of micromeres contributes to gastrulation in the sea urchin embryo."},"authors":{"en":[{"name":"Isaac Shakson"},{"name":"Dubosky Douglas"},{"name":"Waldron Ashley"},{"name":"Emura Natsuko"},{"name":"Furze Aidan"},{"name":"Rao Kavya"},{"name":"Mori Masaru"},{"name":"Ragavendran Ashok"},{"name":"Makinoshima Hideki"},{"name":"Yajima Mamiko"}],"ja":[{"name":"Isaac Shakson"},{"name":"Dubosky Douglas"},{"name":"Waldron Ashley"},{"name":"江村 菜津子"},{"name":"Furze Aidan"},{"name":"Rao Kavya"},{"name":"Mori Masaru"},{"name":"Ragavendran Ashok"},{"name":"Makinoshima Hideki"},{"name":"Yajima Mamiko"}]},"description":{"en":"During development, a group of cells called organizers plays critical roles by sending signals to adjacent cells and controlling embryonic and tissue patterning. Recent studies suggest that these inductive cells facilitate the downstream signaling pathways conserved across organisms. However, what makes these cells fundamentally inductive is little understood. In this study, we demonstrate that the micromeres of the sea urchin, one of the known organizers, have distinct metabolic properties compared to the rest of the embryo. The specific metabolic inhibitors for sugar metabolism (2-DG), fatty acid synthesis (cerulenin), and N-linked glycosylation (tunicamycin) compromise micromeres' regulatory capacity, altering the downstream germ layer patterning in the resultant embryos. Notably, the endoplasmic reticulum (ER) asymmetrically localizes during asymmetric cell division, resulting in the enrichment of ER and Wnt protein at the vegetal cortex of micromeres. Metabolic inhibition appears to compromise ER activity in Wnt particle distribution. We propose that the micromere ER is sensitive to specific metabolic regulation, contributing to the inductive signaling activity. This study provides a paradigm of how ER and metabolic regulation contribute to the inductive capability of the cells.","ja":"During development, a group of cells called organizers plays critical roles by sending signals to adjacent cells and controlling embryonic and tissue patterning. Recent studies suggest that these inductive cells facilitate the downstream signaling pathways conserved across organisms. However, what makes these cells fundamentally inductive is little understood. In this study, we demonstrate that the micromeres of the sea urchin, one of the known organizers, have distinct metabolic properties compared to the rest of the embryo. The specific metabolic inhibitors for sugar metabolism (2-DG), fatty acid synthesis (cerulenin), and N-linked glycosylation (tunicamycin) compromise micromeres' regulatory capacity, altering the downstream germ layer patterning in the resultant embryos. Notably, the endoplasmic reticulum (ER) asymmetrically localizes during asymmetric cell division, resulting in the enrichment of ER and Wnt protein at the vegetal cortex of micromeres. Metabolic inhibition appears to compromise ER activity in Wnt particle distribution. We propose that the micromere ER is sensitive to specific metabolic regulation, contributing to the inductive signaling activity. This study provides a paradigm of how ER and metabolic regulation contribute to the inductive capability of the cells."},"publication_date":"2025-08-11","publication_name":{"en":"Nature Communications","ja":"Nature Communications"},"volume":"16","number":"1","languages":["eng"],"referee":true,"identifiers":{"doi":["10.1038/s41467-025-62697-8"],"issn":["2041-1723"]},"published_paper_type":"scientific_journal"}}
{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/40744298","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467280","label":"url"}],"paper_title":{"en":"Germline factors, TDRD and Piwi, colocalize with Vasa on the mitotic apparatus during the embryogenesis of the sea urchin.","ja":"Germline factors, TDRD and Piwi, colocalize with Vasa on the mitotic apparatus during the embryogenesis of the sea urchin."},"authors":{"en":[{"name":"Witmer Mariana"},{"name":"Mehta Nirali"},{"name":"Emura Natsuko"},{"name":"Yajima Mamiko"}],"ja":[{"name":"Witmer Mariana"},{"name":"Mehta Nirali"},{"name":"江村 菜津子"},{"name":"Yajima Mamiko"}]},"description":{"en":"Germline factors are thought to function exclusively in the germline, providing the unique characteristics of germ cells. However, recent studies suggest that some of these factors may also be expressed and function outside the germline. One such example includes Vasa, a DEAD-box RNA helicase that appears to control localized translation on the spindle, facilitating efficient protein synthesis during embryogenesis of the sea urchin. However, it remains unclear if other germline factors are also involved in this process. In this study, we investigated the localization dynamics of Vasa's partners in the germline, such as Tudor-domain-containing proteins (TDRDs) and P-element-induced wimpy testis proteins (Piwis). Among TDRDs tested in this study, we found that TDRD7 is enriched on the spindle and forms granules with Vasa during early embryogenesis. Vasa and TDRD7 recruited each other when the expression of either was forced at the membrane, suggesting their interaction with each other. TDRD7 mutants lacking the N-terminal eLOTUS domain or the central intrinsically disordered region exhibited reduced granule formation, which also compromised their recruitment to Vasa. In contrast, PiwiL1/2 and PiwiL3 showed enrichment at the perinuclear region and the spindle, yet were never recruited to Vasa or TDRD7 when either was expressed at the membrane. These results suggest that a group of germline factors is present and may dynamically interact with each other on the spindle, contributing to somatic cell regulation in the sea urchin embryo.","ja":"Germline factors are thought to function exclusively in the germline, providing the unique characteristics of germ cells. However, recent studies suggest that some of these factors may also be expressed and function outside the germline. One such example includes Vasa, a DEAD-box RNA helicase that appears to control localized translation on the spindle, facilitating efficient protein synthesis during embryogenesis of the sea urchin. However, it remains unclear if other germline factors are also involved in this process. In this study, we investigated the localization dynamics of Vasa's partners in the germline, such as Tudor-domain-containing proteins (TDRDs) and P-element-induced wimpy testis proteins (Piwis). Among TDRDs tested in this study, we found that TDRD7 is enriched on the spindle and forms granules with Vasa during early embryogenesis. Vasa and TDRD7 recruited each other when the expression of either was forced at the membrane, suggesting their interaction with each other. TDRD7 mutants lacking the N-terminal eLOTUS domain or the central intrinsically disordered region exhibited reduced granule formation, which also compromised their recruitment to Vasa. In contrast, PiwiL1/2 and PiwiL3 showed enrichment at the perinuclear region and the spindle, yet were never recruited to Vasa or TDRD7 when either was expressed at the membrane. These results suggest that a group of germline factors is present and may dynamically interact with each other on the spindle, contributing to somatic cell regulation in the sea urchin embryo."},"publication_date":"2025-07-29","publication_name":{"en":"Developmental Biology","ja":"Developmental Biology"},"volume":"527","starting_page":"98","ending_page":"108","languages":["eng"],"referee":true,"identifiers":{"doi":["10.1016/j.ydbio.2025.07.016"],"issn":["1095-564X"]},"published_paper_type":"scientific_journal"}}
{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/39714020","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467282","label":"url"}],"paper_title":{"en":"The evolutionary modifications of a GoLoco motif in the AGS protein facilitate micromere formation in the sea urchin embryo.","ja":"The evolutionary modifications of a GoLoco motif in the AGS protein facilitate micromere formation in the sea urchin embryo."},"authors":{"en":[{"name":"Emura Natsuko"},{"name":"Wavreil Florence D M"},{"name":"Fries Annaliese"},{"name":"Yajima Mamiko"}],"ja":[{"name":"江村 菜津子"},{"name":"Wavreil Florence D M"},{"name":"Fries Annaliese"},{"name":"Yajima Mamiko"}]},"description":{"en":"The evolutionary introduction of asymmetric cell division (ACD) into the developmental program facilitates the formation of a new cell type, contributing to developmental diversity and, eventually, species diversification. The micromere of the sea urchin embryo may serve as one of those examples: an ACD at the 16-cell stage forms micromeres unique to echinoids among echinoderms. We previously reported that a polarity factor, activator of G-protein signaling (AGS), plays a crucial role in micromere formation. However, AGS and its associated ACD factors are present in all echinoderms and across most metazoans. This raises the question of what evolutionary modifications of AGS protein or its surrounding molecular environment contributed to the evolutionary acquisition of micromeres only in echinoids. In this study, we learned that the GoLoco motifs at the AGS C-terminus play critical roles in regulating micromere formation in sea urchin embryos. Further, other echinoderms' AGS or chimeric AGS that contain the C-terminus of AGS orthologs from various organisms showed varied localization and function in micromere formation. In contrast, the sea star or the pencil urchin orthologs of other ACD factors were consistently localized at the vegetal cortex in the sea urchin embryo, suggesting that AGS may be a unique variable factor that facilitates ACD diversity among echinoderms. Consistently, sea urchin AGS appears to facilitate micromere-like cell formation and accelerate the enrichment timing of the germline factor Vasa during early embryogenesis of the pencil urchin, an ancestral type of sea urchin. Based on these observations, we propose that the molecular evolution of a single polarity factor facilitates ACD diversity while preserving the core ACD machinery among echinoderms and beyond during evolution.","ja":"The evolutionary introduction of asymmetric cell division (ACD) into the developmental program facilitates the formation of a new cell type, contributing to developmental diversity and, eventually, species diversification. The micromere of the sea urchin embryo may serve as one of those examples: an ACD at the 16-cell stage forms micromeres unique to echinoids among echinoderms. We previously reported that a polarity factor, activator of G-protein signaling (AGS), plays a crucial role in micromere formation. However, AGS and its associated ACD factors are present in all echinoderms and across most metazoans. This raises the question of what evolutionary modifications of AGS protein or its surrounding molecular environment contributed to the evolutionary acquisition of micromeres only in echinoids. In this study, we learned that the GoLoco motifs at the AGS C-terminus play critical roles in regulating micromere formation in sea urchin embryos. Further, other echinoderms' AGS or chimeric AGS that contain the C-terminus of AGS orthologs from various organisms showed varied localization and function in micromere formation. In contrast, the sea star or the pencil urchin orthologs of other ACD factors were consistently localized at the vegetal cortex in the sea urchin embryo, suggesting that AGS may be a unique variable factor that facilitates ACD diversity among echinoderms. Consistently, sea urchin AGS appears to facilitate micromere-like cell formation and accelerate the enrichment timing of the germline factor Vasa during early embryogenesis of the pencil urchin, an ancestral type of sea urchin. Based on these observations, we propose that the molecular evolution of a single polarity factor facilitates ACD diversity while preserving the core ACD machinery among echinoderms and beyond during evolution."},"publication_date":"2024-12-23","publication_name":{"en":"eLife","ja":"eLife"},"volume":"13","languages":["eng"],"referee":true,"identifiers":{"doi":["10.7554/eLife.100086"],"issn":["2050-084X"]},"published_paper_type":"scientific_journal"}}
{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/32150685","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467286","label":"url"}],"paper_title":{"en":"Effect of Downregulating the Hippo Pathway Members YAP1 and LATS2 Transcripts on Early Development and Gene Expression Involved in Differentiation in Porcine Embryos.","ja":"Effect of Downregulating the Hippo Pathway Members YAP1 and LATS2 Transcripts on Early Development and Gene Expression Involved in Differentiation in Porcine Embryos."},"authors":{"en":[{"name":"Emura Natsuko"},{"name":"Saito Yuriko"},{"name":"Miura Ruri"},{"name":"Sawai Ken"}],"ja":[{"name":"江村 菜津子"},{"name":"Saito Yuriko"},{"name":"Miura Ruri"},{"name":"Sawai Ken"}]},"description":{"en":"In mouse development, differentiation of the inner cell mass (ICM) and trophectoderm (TE) during the transition from the morula to blastocyst stage is regulated by the Hippo pathway; however, the functions of the Hippo pathway in porcine embryogenesis have not been investigated. In the present study, we examined the gene expression patterns of the Hippo pathway members yes-associated protein 1 (YAP1) and large tumor suppressor 2 (LATS2) and the functions of these genes during porcine preimplantation development using RNA interference. Both YAP1 and LATS2 mRNA levels were shown high in the in vitro matured oocytes and 1-cell stage embryos and fell progressively with development. YAP1 nuclear localization was detected at the morula and blastocyst stages. Downregulation of either YAP1 or LATS2 inhibited porcine preimplantation development and affected the expression levels of POU class 5 homeobox 1 (OCT-4) and SRY-related HMG-box gene 2 (SOX2), transcription factors necessary for the ICM/TE differentiation. Taken together, YAP1 and LATS2 are essential for porcine preimplantation development, and it is possible that the Hippo pathway has important roles in porcine ICM/TE segregation.","ja":"In mouse development, differentiation of the inner cell mass (ICM) and trophectoderm (TE) during the transition from the morula to blastocyst stage is regulated by the Hippo pathway; however, the functions of the Hippo pathway in porcine embryogenesis have not been investigated. In the present study, we examined the gene expression patterns of the Hippo pathway members yes-associated protein 1 (YAP1) and large tumor suppressor 2 (LATS2) and the functions of these genes during porcine preimplantation development using RNA interference. Both YAP1 and LATS2 mRNA levels were shown high in the in vitro matured oocytes and 1-cell stage embryos and fell progressively with development. YAP1 nuclear localization was detected at the morula and blastocyst stages. Downregulation of either YAP1 or LATS2 inhibited porcine preimplantation development and affected the expression levels of POU class 5 homeobox 1 (OCT-4) and SRY-related HMG-box gene 2 (SOX2), transcription factors necessary for the ICM/TE differentiation. Taken together, YAP1 and LATS2 are essential for porcine preimplantation development, and it is possible that the Hippo pathway has important roles in porcine ICM/TE segregation."},"publication_date":"2020-03-05","publication_name":{"en":"Cellular Reprogramming","ja":"Cellular Reprogramming"},"volume":"22","number":"2","starting_page":"62","ending_page":"70","languages":["eng"],"referee":true,"identifiers":{"doi":["10.1089/cell.2019.0082"],"issn":["2152-4998"]},"published_paper_type":"scientific_journal"}}
{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/32093223","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467283","label":"url"}],"paper_title":{"en":"Steroidogenic Factor 1 (NR5A1) Activates ATF3 Transcriptional Activity.","ja":"Steroidogenic Factor 1 (NR5A1) Activates ATF3 Transcriptional Activity."},"authors":{"en":[{"name":"Emura Natsuko"},{"name":"Wang Chiung-Min"},{"name":"Yang William Harry"},{"name":"Yang Wei-Hsiung"}],"ja":[{"name":"江村 菜津子"},{"name":"Wang Chiung-Min"},{"name":"Yang William Harry"},{"name":"Yang Wei-Hsiung"}]},"description":{"en":"Steroidogenic Factor 1 (SF-1/NR5A1), an orphan nuclear receptor, is important for sexual differentiation and the development of multiple endocrine organs, as well as cell proliferation in cancer cells. Activating transcription factor 3 (ATF3) is a transcriptional repressor, and its expression is rapidly induced by DNA damage and oncogenic stimuli. Since both NR5A1 and ATF3 can regulate and cooperate with several transcription factors, we hypothesized that NR5A1 may interact with ATF3 and plays a functional role in cancer development. First, we found that NR5A1 physically interacts with ATF3. We further demonstrated that ATF3 expression is up-regulated by NR5A1. Moreover, the promoter activity of the ATF3 is activated by NR5A1 in a dose-dependent manner in several cell lines. By mapping the ATF3 promoter as well as the site-directed mutagenesis analysis, we provide evidence that NR5A1 response elements (-695 bp and -665 bp) are required for ATF3 expression by NR5A1. It is well known that the transcriptional activities of NR5A1 are modulated by post-translational modifications, such as small ubiquitin-related modifier (SUMO) modification and phosphorylation. Notably, we found that both SUMOylation and phosphorylation of NR5A1 play roles, at least in part, for NR5A1-mediated ATF3 expression. Overall, our results provide the first evidence of a novel relationship between NR5A1 and ATF3.","ja":"Steroidogenic Factor 1 (SF-1/NR5A1), an orphan nuclear receptor, is important for sexual differentiation and the development of multiple endocrine organs, as well as cell proliferation in cancer cells. Activating transcription factor 3 (ATF3) is a transcriptional repressor, and its expression is rapidly induced by DNA damage and oncogenic stimuli. Since both NR5A1 and ATF3 can regulate and cooperate with several transcription factors, we hypothesized that NR5A1 may interact with ATF3 and plays a functional role in cancer development. First, we found that NR5A1 physically interacts with ATF3. We further demonstrated that ATF3 expression is up-regulated by NR5A1. Moreover, the promoter activity of the ATF3 is activated by NR5A1 in a dose-dependent manner in several cell lines. By mapping the ATF3 promoter as well as the site-directed mutagenesis analysis, we provide evidence that NR5A1 response elements (-695 bp and -665 bp) are required for ATF3 expression by NR5A1. It is well known that the transcriptional activities of NR5A1 are modulated by post-translational modifications, such as small ubiquitin-related modifier (SUMO) modification and phosphorylation. Notably, we found that both SUMOylation and phosphorylation of NR5A1 play roles, at least in part, for NR5A1-mediated ATF3 expression. Overall, our results provide the first evidence of a novel relationship between NR5A1 and ATF3."},"publication_date":"2020-02-20","publication_name":{"en":"International Journal of Molecular Sciences","ja":"International Journal of Molecular Sciences"},"volume":"21","number":"4","languages":["eng"],"referee":true,"identifiers":{"doi":["10.3390/ijms21041429"],"issn":["1422-0067"]},"published_paper_type":"scientific_journal"}}
{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/31130592","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467288","label":"url"}],"paper_title":{"en":"The necessity of TEAD4 for early development and gene expression involved in differentiation in porcine embryos.","ja":"The necessity of TEAD4 for early development and gene expression involved in differentiation in porcine embryos."},"authors":{"en":[{"name":"Emura Natsuko"},{"name":"Takahashi Kazuki"},{"name":"Saito Yuriko"},{"name":"Sawai Ken"}],"ja":[{"name":"江村 菜津子"},{"name":"Takahashi Kazuki"},{"name":"Saito Yuriko"},{"name":"Sawai Ken"}]},"description":{"en":"TEA domain family transcription factor 4 (Tead4) is known to be important for the trophectoderm (TE) segregation in murine embryos. However, the role of TEAD4 in early development of porcine embryos is still unknown. We examined TEAD4 expression patterns and attempted to determine the functions of TEAD4 during porcine preimplantation development using RNA interference. TEAD4 mRNA was upregulated from the 2-4-cell to 8-16-cell stages and then decreased to the blastocyst stage. Nuclear localization of TEAD4 protein was detected at the 16-cell stage, as well as at subsequent developmental stages. In porcine embryos injected with TEAD4 siRNA, transformation from morula to blastocyst was inhibited. Although TEAD4 downregulation did not affect the expression levels of POU class 5 homeobox 1 (OCT-4), transcription of SRY-related HMG-box gene 2 (SOX2) was detected at high level in TEAD4-downregulated embryos. It is possible that TEAD4 contributes to blastocyst formation in porcine embryos through downregulation of SOX2 expression. Collectively, our results indicate that TEAD4 is an important factor for the preimplantation development of porcine embryos.","ja":"TEA domain family transcription factor 4 (Tead4) is known to be important for the trophectoderm (TE) segregation in murine embryos. However, the role of TEAD4 in early development of porcine embryos is still unknown. We examined TEAD4 expression patterns and attempted to determine the functions of TEAD4 during porcine preimplantation development using RNA interference. TEAD4 mRNA was upregulated from the 2-4-cell to 8-16-cell stages and then decreased to the blastocyst stage. Nuclear localization of TEAD4 protein was detected at the 16-cell stage, as well as at subsequent developmental stages. In porcine embryos injected with TEAD4 siRNA, transformation from morula to blastocyst was inhibited. Although TEAD4 downregulation did not affect the expression levels of POU class 5 homeobox 1 (OCT-4), transcription of SRY-related HMG-box gene 2 (SOX2) was detected at high level in TEAD4-downregulated embryos. It is possible that TEAD4 contributes to blastocyst formation in porcine embryos through downregulation of SOX2 expression. Collectively, our results indicate that TEAD4 is an important factor for the preimplantation development of porcine embryos."},"publication_date":"2019-05-25","publication_name":{"en":"The Journal of Reproduction and Development","ja":"The Journal of Reproduction and Development"},"volume":"65","number":"4","starting_page":"361","ending_page":"368","languages":["eng"],"referee":true,"identifiers":{"doi":["10.1262/jrd.2018-120"],"issn":["1348-4400"]},"published_paper_type":"scientific_journal"}}
{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/27941302","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467287","label":"url"}],"paper_title":{"en":"Effects of downregulating TEAD4 transcripts by RNA interference on early development of bovine embryos.","ja":"Effects of downregulating TEAD4 transcripts by RNA interference on early development of bovine embryos."},"authors":{"en":[{"name":"Sakurai Nobuyuki"},{"name":"Takahashi Kazuki"},{"name":"Emura Natsuko"},{"name":"Hashizume Tsutomu"},{"name":"Sawai Ken"}],"ja":[{"name":"Sakurai Nobuyuki"},{"name":"Takahashi Kazuki"},{"name":"江村 菜津子"},{"name":"Hashizume Tsutomu"},{"name":"Sawai Ken"}]},"description":{"en":"Transcription factor TEA domain family transcription factor 4 (Tead4) is one of the key factors involved in the differentiation of the trophectoderm (TE) in murine embryos. However, knowledge on the roles of TEAD4 in preimplantation development during bovine embryos is currently limited. This study examined the transcript and protein expression patterns of TEAD4 and attempted to elucidate the functions of TEAD4 during bovine preimplantation development using RNA interference. TEAD4 mRNA was found to be upregulated between the 16-cell and morula stages, and nuclear localization of the TEAD4 protein was detected at the morula stage, as well as in subsequent developmental stages. TEAD4 downregulation did not affect embryonic development until the blastocyst stage, and TEAD4-downregulated embryos were capable of forming the TE under both 5% and 21% O2 conditions. Results of gene expression analysis showed that TEAD4 downregulation did not affect the expression levels of POU class 5 transcription factor 1 (OCT-4), NANOG, caudal-type homeobox 2 (CDX2), GATA binding protein 3 (GATA3), and interferon-tau (IFNT). In conclusion, TEAD4 might be dispensable for development until the blastocyst stage and TE differentiation in bovine embryos.","ja":"Transcription factor TEA domain family transcription factor 4 (Tead4) is one of the key factors involved in the differentiation of the trophectoderm (TE) in murine embryos. However, knowledge on the roles of TEAD4 in preimplantation development during bovine embryos is currently limited. This study examined the transcript and protein expression patterns of TEAD4 and attempted to elucidate the functions of TEAD4 during bovine preimplantation development using RNA interference. TEAD4 mRNA was found to be upregulated between the 16-cell and morula stages, and nuclear localization of the TEAD4 protein was detected at the morula stage, as well as in subsequent developmental stages. TEAD4 downregulation did not affect embryonic development until the blastocyst stage, and TEAD4-downregulated embryos were capable of forming the TE under both 5% and 21% O2 conditions. Results of gene expression analysis showed that TEAD4 downregulation did not affect the expression levels of POU class 5 transcription factor 1 (OCT-4), NANOG, caudal-type homeobox 2 (CDX2), GATA binding protein 3 (GATA3), and interferon-tau (IFNT). In conclusion, TEAD4 might be dispensable for development until the blastocyst stage and TE differentiation in bovine embryos."},"publication_date":"2016-12-11","publication_name":{"en":"The Journal of Reproduction and Development","ja":"The Journal of Reproduction and Development"},"volume":"63","number":"2","starting_page":"135","ending_page":"142","languages":["eng"],"referee":true,"identifiers":{"doi":["10.1262/jrd.2016-130"],"issn":["1348-4400"]},"published_paper_type":"scientific_journal"}}
{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/27500421","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467289","label":"url"}],"paper_title":{"en":"The Necessity of OCT-4 and CDX2 for Early Development and Gene Expression Involved in Differentiation of Inner Cell Mass and Trophectoderm Lineages in Bovine Embryos.","ja":"The Necessity of OCT-4 and CDX2 for Early Development and Gene Expression Involved in Differentiation of Inner Cell Mass and Trophectoderm Lineages in Bovine Embryos."},"authors":{"en":[{"name":"Sakurai Nobuyuki"},{"name":"Takahashi Kazuki"},{"name":"Emura Natsuko"},{"name":"Fujii Takashi"},{"name":"Hirayama Hiroki"},{"name":"Kageyama Soichi"},{"name":"Hashizume Tsutomu"},{"name":"Sawai Ken"}],"ja":[{"name":"Sakurai Nobuyuki"},{"name":"Takahashi Kazuki"},{"name":"江村 菜津子"},{"name":"Fujii Takashi"},{"name":"Hirayama Hiroki"},{"name":"Kageyama Soichi"},{"name":"Hashizume Tsutomu"},{"name":"Sawai Ken"}]},"description":{"en":"The functions of POU class 5 transcription factor 1 (Oct-4) and caudal-type homeobox 2 (Cdx2) in the differentiation of the murine inner cell mass (ICM) and trophectoderm (TE) have been described in detail. However, little is known about the roles of OCT-4 and CDX2 in preimplantation bovine embryos. To elucidate their functions during early development in bovine embryos, we performed OCT-4 and CDX2 downregulation using RNA interference. We injected OCT-4- or CDX2-specific short interfering RNAs (siRNAs) into bovine zygotes. The rate of blastocyst development of OCT-4-downregulated embryos was lower compared with uninjected or control siRNA-injected embryos. Gene expression analysis revealed decreased CDX2 and fibroblast growth factor 4 expression in OCT-4-downregulated embryos. CDX2-downregulated embryos developed to the blastocyst stage; however, in most cases, blastocoel formation was delayed. Gene expression analysis revealed decreased GATA3 expression and elevated NANOG expression in CDX2-downregulated embryos. In conclusion, OCT-4 and CDX2 are essential for early development and gene expression involved in differentiation of ICM and TE lineages in bovine embryos.","ja":"The functions of POU class 5 transcription factor 1 (Oct-4) and caudal-type homeobox 2 (Cdx2) in the differentiation of the murine inner cell mass (ICM) and trophectoderm (TE) have been described in detail. However, little is known about the roles of OCT-4 and CDX2 in preimplantation bovine embryos. To elucidate their functions during early development in bovine embryos, we performed OCT-4 and CDX2 downregulation using RNA interference. We injected OCT-4- or CDX2-specific short interfering RNAs (siRNAs) into bovine zygotes. The rate of blastocyst development of OCT-4-downregulated embryos was lower compared with uninjected or control siRNA-injected embryos. Gene expression analysis revealed decreased CDX2 and fibroblast growth factor 4 expression in OCT-4-downregulated embryos. CDX2-downregulated embryos developed to the blastocyst stage; however, in most cases, blastocoel formation was delayed. Gene expression analysis revealed decreased GATA3 expression and elevated NANOG expression in CDX2-downregulated embryos. In conclusion, OCT-4 and CDX2 are essential for early development and gene expression involved in differentiation of ICM and TE lineages in bovine embryos."},"publication_date":"2016-08-08","publication_name":{"en":"Cellular Reprogramming","ja":"Cellular Reprogramming"},"volume":"18","number":"5","starting_page":"309","ending_page":"318","languages":["eng"],"referee":true,"identifiers":{"doi":["10.1089/cell.2015.0081"],"issn":["2152-4998"]},"published_paper_type":"scientific_journal"}}
{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/27210587","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467291","label":"url"}],"paper_title":{"en":"OCT-4 expression is essential for the segregation of trophectoderm lineages in porcine preimplantation embryos.","ja":"OCT-4 expression is essential for the segregation of trophectoderm lineages in porcine preimplantation embryos."},"authors":{"en":[{"name":"Emura Natsuko"},{"name":"Sakurai Nobuyuki"},{"name":"Takahashi Kazuki"},{"name":"Hashizume Tsutomu"},{"name":"Sawai Ken"}],"ja":[{"name":"江村 菜津子"},{"name":"Sakurai Nobuyuki"},{"name":"Takahashi Kazuki"},{"name":"Hashizume Tsutomu"},{"name":"Sawai Ken"}]},"description":{"en":"Oct-4, a member of the POU family of transcription factors, is a key factor that regulates the segregation of the inner cell mass (ICM) and the trophectoderm (TE) during the transition from morula to blastocyst in mice. However, little is known about its role in porcine early embryogenesis. To determine the function of OCT-4 in the ICM and TE segregation of porcine embryos, we studied the developmental morphology of porcine embryos using RNA interference technology. Our experiments demonstrated that when 1-cell stage embryos were co-injected with the small interfering RNA (siRNA)for targeted knockdown of OCT-4 (OCT-4-siRNA) and tetramethylrhodamine isothiocyanate (TRITC)-dextran conjugate (Dx), they failed to form blastocysts. Therefore, in this study, we constructed chimeric embryos comprising blastomeres that either expressed OCT-4 normally or showed downregulated OCT-4 expression by co-injection of OCT-4-siRNA and Dx into one blastomere in 2- to 4-cell stage embryos. In control embryos, which were co-injected with control siRNA and Dx, Dx-positive cells contributed to the TE lineage in almost all the blastocysts examined. In contrast, Dx-positive cells derived from a blastomere co-injected with OCT-4-siRNA and Dx were degenerated in almost half the blastocysts. This was probably due to the inability of these cells to differentiate into the TE lineage. Real-time RT-PCR analysis revealed no difference in the levels of SOX2, TEAD4, FGF4 and FGFR1-IIIc, all of which are known to be regulated by OCT-4, between the OCT-4-siRNA-injected morulae and the control ones. However, the level of CDX2, a molecule specifically expressed in the TE lineage, was significantly higher in the former than in the latter. Our results indicate that continuous expression of OCT-4 in blastomeres is essential for TE formation of porcine embryos.","ja":"Oct-4, a member of the POU family of transcription factors, is a key factor that regulates the segregation of the inner cell mass (ICM) and the trophectoderm (TE) during the transition from morula to blastocyst in mice. However, little is known about its role in porcine early embryogenesis. To determine the function of OCT-4 in the ICM and TE segregation of porcine embryos, we studied the developmental morphology of porcine embryos using RNA interference technology. Our experiments demonstrated that when 1-cell stage embryos were co-injected with the small interfering RNA (siRNA)for targeted knockdown of OCT-4 (OCT-4-siRNA) and tetramethylrhodamine isothiocyanate (TRITC)-dextran conjugate (Dx), they failed to form blastocysts. Therefore, in this study, we constructed chimeric embryos comprising blastomeres that either expressed OCT-4 normally or showed downregulated OCT-4 expression by co-injection of OCT-4-siRNA and Dx into one blastomere in 2- to 4-cell stage embryos. In control embryos, which were co-injected with control siRNA and Dx, Dx-positive cells contributed to the TE lineage in almost all the blastocysts examined. In contrast, Dx-positive cells derived from a blastomere co-injected with OCT-4-siRNA and Dx were degenerated in almost half the blastocysts. This was probably due to the inability of these cells to differentiate into the TE lineage. Real-time RT-PCR analysis revealed no difference in the levels of SOX2, TEAD4, FGF4 and FGFR1-IIIc, all of which are known to be regulated by OCT-4, between the OCT-4-siRNA-injected morulae and the control ones. However, the level of CDX2, a molecule specifically expressed in the TE lineage, was significantly higher in the former than in the latter. Our results indicate that continuous expression of OCT-4 in blastomeres is essential for TE formation of porcine embryos."},"publication_date":"2016-05-20","publication_name":{"en":"The Journal of Reproduction and Development","ja":"The Journal of Reproduction and Development"},"volume":"62","number":"4","starting_page":"401","ending_page":"408","languages":["eng"],"referee":true,"identifiers":{"doi":["10.1262/jrd.2016-040"],"issn":["1348-4400"]},"published_paper_type":"scientific_journal"}}
{"insert":{"user_id":"R000059838","type":"published_papers"},"force":{"see_also":[{"@id":"https://www.ncbi.nlm.nih.gov/pubmed/26074126","label":"url"},{"@id":"https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=467284","label":"url"}],"paper_title":{"en":"Effects of downregulating GLIS1 transcript on preimplantation development and gene expression of bovine embryos.","ja":"Effects of downregulating GLIS1 transcript on preimplantation development and gene expression of bovine embryos."},"authors":{"en":[{"name":"Takahashi Kazuki"},{"name":"Sakurai Nobuyuki"},{"name":"Emura Natsuko"},{"name":"Hashizume Tsutomu"},{"name":"Sawai Ken"}],"ja":[{"name":"Takahashi Kazuki"},{"name":"Sakurai Nobuyuki"},{"name":"江村 菜津子"},{"name":"Hashizume Tsutomu"},{"name":"Sawai Ken"}]},"description":{"en":"Krüppel-like protein Gli-similar 1 (GLIS1) is known as a direct reprogramming factor for the generation of induced pluripotent stem cells. The objective of this study was to investigate the role of GLIS1 in the preimplantation development of bovine embryos. GLIS1 transcripts in in vitro-matured oocytes and 1-cell to 4-cell stage embryos were detected, but they were either absent or at trace levels at the 8-cell to blastocyst stages. We attempted GLIS1 downregulation of bovine early embryos by RNA interference and evaluated developmental competency and gene transcripts, which are involved in zygotic gene activation (ZGA) in GLIS1-downregulated embryos. Injection of specific siRNA resulted in a distinct decrease in GLIS1 transcript in bovine embryos at the 4-cell stage. Although the bovine embryos injected with GLIS1-siRNA could develop to the 16-cell stage, these embryos had difficulty in developing beyond the 32-cell stage. Gene transcripts of PDHA1 and HSPA8, which are transcribed after ZGA, showed lower level in GLIS1 downregulated embryos. It is possible that GLIS1-downregulated embryos fail to initiate ZGA. Our results indicated that GLIS1 is an important factor for the preimplantation development of bovine embryos.","ja":"Krüppel-like protein Gli-similar 1 (GLIS1) is known as a direct reprogramming factor for the generation of induced pluripotent stem cells. The objective of this study was to investigate the role of GLIS1 in the preimplantation development of bovine embryos. GLIS1 transcripts in in vitro-matured oocytes and 1-cell to 4-cell stage embryos were detected, but they were either absent or at trace levels at the 8-cell to blastocyst stages. We attempted GLIS1 downregulation of bovine early embryos by RNA interference and evaluated developmental competency and gene transcripts, which are involved in zygotic gene activation (ZGA) in GLIS1-downregulated embryos. Injection of specific siRNA resulted in a distinct decrease in GLIS1 transcript in bovine embryos at the 4-cell stage. Although the bovine embryos injected with GLIS1-siRNA could develop to the 16-cell stage, these embryos had difficulty in developing beyond the 32-cell stage. Gene transcripts of PDHA1 and HSPA8, which are transcribed after ZGA, showed lower level in GLIS1 downregulated embryos. It is possible that GLIS1-downregulated embryos fail to initiate ZGA. Our results indicated that GLIS1 is an important factor for the preimplantation development of bovine embryos."},"publication_date":"2015-06-15","publication_name":{"en":"The Journal of Reproduction and Development","ja":"The Journal of Reproduction and Development"},"volume":"61","number":"5","starting_page":"369","ending_page":"374","languages":["eng"],"referee":true,"identifiers":{"doi":["10.1262/jrd.2015-029"],"issn":["1348-4400"]},"published_paper_type":"scientific_journal"}}