Takekazu Horio, Jinpei Yamashita, Katsuzo Nishikawa, Tomisaburo Kakuno, Kazuo Hosoi, Junnosuke Suzuki and Setsuko Yoshimura : Organization of energy transducing membranes, University of Tokyo Press, Tokyo, Aug. 1973.
学術論文(審査論文):
1.
Jing Tang, Chenjuan Yao, Yingqi Liu, Jiaming Yuan, Li Wu, Kazuo Hosoi, Shali Yu, Chunyan Huang, Haiyan Wei and Gang Chen : Arsenic trioxide induces expression of BCL-2 expression via NF-κB and p38 MAPK signaling pathways in BEAS-2B cells during apoptosis, Ecotoxicology and Environmental Safety, 222, 112531, 2021.
(要約)
Inorganic arsenic compounds are environmental toxicants that are widely distributed in air, water, and food. B-cell lymphoma 2 (BCL-2) is an oncogene having anti-apoptotic function. In this study, we clarify that BCL-2, as a pro-apoptotic factor, participates in AsO-induced apoptosis in BEAS-2B cells. Specifically, AsO stimulated the expression of BCL-2 mRNA and protein in a dose-dependent manner which was highly accumulated in the nucleus of BEAS-2B cell together with chromatin condensation and DNA fragmentation during apoptosis. Mechanistically, the process described above is mediated through the NF-κB and p38 MAPK signaling pathways, which can be abated by corresponding inhibitors, such as BAY11-7082 and SB203580, respectively. Additionally, BAY11-7082, actinomycin D, and cycloheximide have inhibitory effects on AsO-induced expression of BCL-2 mRNA and protein, and restore the cell viability of BEAS-2B cells. Suppression of BCL-2 protein activation by ABT-199 also restored viability of BEAS-2B cell in AsO-induced apoptosis. Furthermore, AsO increased the level of BCL-2 phosphorylation. These results suggest that in BEAS-2B cells, AsO-induced apoptosis is mainly dominated by BCL-2 upregulation, nuclear localization and phosphorylation. The study presented here provides a novel insight into the molecular mechanism of BCL-2-induced apoptosis.
Yingqi Liu, Haiyan Wei, Jing Tang, Jiaming Yuan, Mingmin Wu, Chenjuan Yao, Kazuo Hosoi, Shali Yu, Xinyuan Zhao, Yu Han and Gang Chen : Dysfunction of pulmonary epithelial tight junction induced by silicon dioxide nanoparticles via the ROS/ERK pathway and protein degradation., Chemosphere, 255, 126954, 2020.
(要約)
Silica nanoparticles (SiNPs) are one of the most widely used types of nanoparticles across many industrial sectors, and are known to be present in the air year-round. In this study, we aimed to evaluate the potential adverse effects of SiNP exposure on pulmonary epithelial tight junctions, which serve as a critical barrier between the respiratory system and the circulatory system. In vivo studies confirmed that SiNPs decreased the protein expression levels of zonula occludens 1 (ZO-1), zonula occludens 2 (ZO-2), and occludin in the lungs of C57BL/6 mice. In vitro studies showed that SiNPs not only decreased the mRNA and protein expression of ZO-1 and ZO-2, but also decreased the protein expression of occludin in human bronchial epithelial (BEAS-2B) cells. In addition, SiNP exposure increased reactive oxygen species (ROS) production and activated extracellular regulated protein kinases (ERKs) and c-Jun N-terminal kinase (JNK). The inhibition of ROS and ERKs effectively protected the SiNP-induced downregulation of ZO-1 mRNA and protein expression, but had no effect on ZO-2 or occludin expression. SiNP-induced matrix metalloproteinase 9 (MMP9) protein expression appeared to be involved in occludin proteolytic degradation, in addition to SiNP-induced direct occludin protein degradation. The present study suggests that SiNPs disturb pulmonary epithelial tight junction structure and function via the ROS/ERK pathway and protein degradation.
Javkhlan Purevjav, Xu Guangfei, Chen Gang, Chenjuan Yao, Yuka Hiroshima, Hiroshi Yoshimura, Toshihiko Nagata and Kazuo Hosoi : Expression and LPS-Induced Elevation of Nod2 and Calprotectin in the Submandibular Gland of Wild-Type and TLR4-Knockout Male Mice, Journal of Research and Practice in Dentistry, 2015, 290259, 2015.
Gang Chen, Chenjuan Yao, Takahiro Hasegawa, Tetsuya Akamatsu, Hiroshi Yoshimura and Kazuo Hosoi : Effects of isoproterenol on aquaporin 5 levels in the parotid gland of mice in vivo., American Journal of Physiology, Endocrinology and Metabolism, 306, 1, E100-E108, 2014.
(要約)
In the membrane fraction of mouse parotid gland (PG), the protein level of aquaporin 5 (AQP5), a member of the water channel family, was increased by injection (ip) of isoproterenol (IPR), a β-adrenergic agonist, at 1 h, and stayed at high levels until 6 h; this change occurred simultaneously as amylase secretion. The AQP5 level then decreased and returned toward the original level at 12-48 h. After IPR injection, the AQP5 mRNA gradually increased and reached a maximum at 24 h. The facts suggest a rapid appearance of AQP5 at plasma membrane by IPR and subsequent degradation/metabolism by activation of proteolytic systems. Pretreatment of animals with two calpain inhibitors, N-Ac-Leu-Leu-methininal (ALLM) and calpeptin, as well as a protein synthesis inhibitor, cycloheximide (CHX), significantly suppressed the IPR-induced AQP5 degradation in the PG membrane fraction; such suppression was not observed by two proteasome inhibitors, MG132 and lactacystin, or the lysosome denaturant chloroquine, although most of these inhibitors increased AQP5 protein levels in unstimulated mice. The AQP5 protein was also degraded by μ-calpain in vitro. Furthermore, we demonstrated that μ-calpain was colocalized with AQP5 in the acinar cells by immunohistochemistry, and its activity in the PG was increased at 6 h after IPR injection. These results suggest that the calpain system was responsible for IPR-induced AQP5 degradation in the parotid gland and that such a system was coupled to the secretory-restoration cycle of amylase in the PG.
Keitaro Satoh, Yoshiteru Seo, Shinsuke Matsuo, Mileva Ratko Karabasil, Miwako Matsuki-Fukushima, Takashi Nakahari and Kazuo Hosoi : Roles of AQP5/AQP5-G103D in carbamylcholine-induced volume decrease and in reduction of the activation energy for water transport by rat parotid acinar cells, Pflügers Archiv : European Journal of Physiology, 464, 4, 375-389, 2012.
(要約)
In order to assess the contribution of the water channel aquaporin-5 (AQP5) to water transport by salivary gland acinar cells, we measured the cell volume and activation energy (E (a)) of diffusive water permeability in isolated parotid acinar cells obtained from AQP5-G103D mutant and their wild-type rats. Immunohistochemistry showed that there was no change induced by carbamylcholine (CCh; 1 μM) in the AQP5 detected in the acinar cells in the wild-type rat. Acinar cells from mutant rats, producing low levels of AQP5 in the apical membrane, showed a minimal increase in the AQP5 due to the CCh. In the wild-type rat, CCh caused a transient swelling of the acinus, followed by a rapid agonist-induced cell shrinkage, reaching a plateau at 30 s. In the mutant rat, the acinus did not swell by CCh challenge, and the agonist-induced cell shrinkage was delayed by 8 s, reaching a transient minimum at around 1 min, and recovered spontaneously even though CCh was persistently present. In the unstimulated wild-type acinar cells, E (a) was 3.4 ± 0.6 kcal mol(-1) and showed no detectable change after CCh stimulation. In the unstimulated mutant acinar cells, high E (a) value (5.9 ± 0.1 kcal mol(-1)) was detected and showed a minimal decrease after CCh stimulation (5.0 ± 0.3 kcal mol(-1)). These results suggested that AQP5 was the main pathway for water transport in the acinar cells and that it was responsible for the rapid agonist-induced acinar cell shrinkage and also necessary to keep the acinar cell volume reduced during the steady secretion in the wild-type rat.
Nunuk Purwanti, Mileva Ratko Karabasil, Shinsuke Matsuo, Gang Chen, Javkhlan Purevjav, Ahmad Azlina, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : Induction of Sca-1 via activation of STAT3 system in the duct cells of the mouse submandibular gland by ligation of the main excretory duct, American Journal of Physiology, Gastrointestinal and Liver Physiology, 301, 5, G814-G824, 2011.
(要約)
To examine the very initial step that takes place immediately after tissue injury and is linked to tissue regeneration, we employed the submandibular gland (SMG), which was injured by ligation of its main excretory duct (MED). Ligation of the MED of the SMG in mice induced the expression of Sca-1, a protein marker of hematopoietic stem cells. In the normal gland, a low level of Sca-1 was expressed, which was localized predominantly in the excretory duct cells. At 1 day after ligation, Sca-1 expression increased prominently in almost all of cells in the duct system, but not in the acinar cells. The level of Sca-1 mRNA had begun to increase at 6 h after ligation and continuously rose thereafter until it reached a plateau, which occurred ∼12 h after ligation. STAT3 phosphorylated at its tyrosine-705 (p-STAT3) in the ligated gland increased immediately after ligation, and it was localized in the nuclei of all duct cells. The results of an EMSA revealed the specific binding of a nuclear extract to the sequence of the γ-interferon activation site (GAS) present in the Sca-1 promoter and confirmed that such binding increased after ligation. Thus the present study suggests that STAT3, having been phosphorylated following MED ligation, was transferred to the nucleus, where it bound to the GAS element in the promoter of Sca-1 gene, resulting in promotion of Sca-1 gene expression. Actual prevention of STAT3 phosphorylation reduced the ligation-induced Sca-1 elevation.
Purevjav Javkhlan, Yuka Hiroshima, Ahmad Azlina, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu, Jun-ichi Kido, Toshihiko Nagata and Kazuo Hosoi : Lipopolysaccharide-mediated induction of calprotectin in the submandibular and parotid glands of mice, Inflammation, 34, 6, 668-680, 2011.
(要約)
S100A8 and S100A9 constitute a heterodimeric protein, calprotectin. The mRNAs of S100A8 and S100A9, being expressed at minimal levels in the submandibular and parotid glands (SMG and PG, respectively) of C3H/HeN mice, were induced strongly and transiently by lipopolysaccharide (LPS). Among the mRNAs of members of the S100 protein family examined, those of S100A8 and S100A9 were specifically induced by LPS in the salivary glands. The induction was assumed to be mediated via toll-like receptor 4 (TLR4), since their elevation was limited in C3H/HeJ mice, a TLR4-mutant strain. These proteins became expressed in the granular convoluted tubular cells and striated duct cells in the SMG, and in both acinar and duct cells in the PG (all in the cytoplasm). The salivary calprotectin level was not increased by LPS treatment, implying that elevated calprotectin was not secreted into the saliva and that they may function in microcellular environment of the salivary gland.
Takahiro Hasegawa, Ahmad Azlina, Javkhlan Purevjav, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : Novel phosphorylation of aquaporin-5 at its threonine 259 through cAMP signaling, American Journal of Physiology, Cell Physiology, 301, 3, C667-C678, 2011.
(要約)
Aquaporin-5 (AQP5), a water channel, plays key roles in salivary secretion. The novel phosphorylation of AQP5 was investigated by using human salivary gland (HSG) cells and mouse salivary glands. In the HSG cells stably transfected with a wild-type mouse AQP5 construct, a protein band immunoreactive with antibody against phosphorylated PKA substrate was detected in the AQP5 immunoprecipitated sample, and its intensity was enhanced by short-term treatment of the cells with 8-bromo-cAMP, forskolin, or phorbol 12-myristate 13-acetate, but not by that with A23187 calcium ionophore. Such enhancement was inhibited in the presence of H-89, a PKA inhibitor. An AQP5 mutant (AQP5-T259A) expressed by transfection of HSG cells was not recognized by anti-phosphorylated PKA substrate antibody, even when the cells were stimulated with the protein kinase activators. Immunoblotting and immunofluorescence studies using a specific antibody detecting AQP5 phosphorylated at its Thr259 demonstrated that AQP5 was rapidly and transiently phosphorylated at the apical membrane of acinar cells in the submandibular and parotid glands after administration of isoproterenol, but not pilocarpine. Furthermore, both AQP5 and AQP5-T259A were constitutively localized at the plasma membrane in HSG cells under the resting and forskolin-stimulated conditions. These results suggest that AQP5 is phosphorylated at its Thr259 by PKA through cAMP, but not Ca(2+), signaling pathways, and that this phosphorylation does not contribute to AQP5 trafficking in the salivary gland cells.
Nunuk Purwanti, Daisuke Tsuji, Ahmad Azlina, Mileva Ratko Karabasil, Purevjav Javkhlan, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu, Kouji Itou and Kazuo Hosoi : Induction of Sca-1 in the duct cells of the mouse submandibular gland by obstruction of the main excretory duct, Journal of Oral Pathology & Medicine, 40, 8, 651-658, 2011.
(要約)
The effect of ligation of the main excretory duct (MED) of the mouse submandibular gland (SMG) on the expression of Sca-1, a stem cell antigen, was examined by Western blotting and immunohistochemistry. By Western blotting, the expression of Sca-1 with a molecular weight of 18 kDa was identified in the normal gland. At 1 day post-ligation, the expression level of Sca-1 was strongly increased in the experimental gland and weakly in the contralateral gland, and such expression in both glands decreased at 6 days. By immunohistochemistry, Sca-1 was detected weakly in the apical membrane of excretory duct (ED) cells of the SMG under the normal condition. By duct ligation, Sca-1 became expressed strongly in most cells of the two major duct systems, i.e., the striated duct (SD) and granular convoluted tubules (GCT), but was not detected in the acinar (Ac) cells. By fluorescence-activated cell sorter (FACS) analysis, the number of side population (SP) cells in this gland was found to be increased by ligation. These results imply that Sca-1-positive cells may have a role in the duct cell proliferation in the regeneration step elicited by MED ligation-induced injury.
Yuka Hiroshima, Mika Bandou, Masatoshi Kataoka, Yuji Inagaki, Mark C. Herzberg, Karen F. Ross, Kazuo Hosoi, Toshihiko Nagata and Jun-ichi Kido : Regulation of antimicrobial peptide expression in human gingival keratinocytes by interleukin-1α, Archives of Oral Biology, 56, 8, 761-767, 2011.
(要約)
In the oral cavity, mucosal keratinocytes resist bacterial infection, in part, by producing broad-spectrum antimicrobial peptides (AMPs) including defensin, adrenomedullin and calprotectin. Epidermal keratinocyte expression of many AMPs increases in response to interleukin-1α (IL-1α). IL-1α is produced by epidermal keratinocytes and regulates cell differentiation. To better understand innate immunity in the oral cavity, we sought to determine how IL-1α might regulate expression of AMPs by human gingival keratinocytes (HGKs) using DNA microarray and Western blot analyses. HGKs from three subjects expressed eleven AMPs, including S100A7, S100A8, S100A9, S100A12, secretory leucocyte protease inhibitor, lipocalin 2 (LCN2), cystatin C and β-defensin 2. Of the expressed AMPs, S100A7, S100A12 and LCN2 were up-regulated by IL-1α (inducible AMPs); the other AMPs were considered to be constitutive. Human gingival keratinocytes, therefore, express constitutive and IL-1α-inducible AMPs to provide a rapid and robust innate response to microbial infection.
Mileva Ratko Karabasil, Takahiro Hasegawa, Ahmad Azlina, Nunuk Purwanti, Chenjuan Yao, Tetsuya Akamatsu, Shigemasa Tomioka and Kazuo Hosoi : Effects of naturally occurring G103D point mutation of AQP5 on its water permeability, trafficking, and cellular localization in the submandibular gland of rats, Biology of the Cell, 103, 2, 69-86, 2011.
(要約)
AQPs (aquaporins) are water channel proteins that are expressed in almost all living things. In mammalians, 13 members of AQPs (AQP0-12) have been identified so far. AQP5 is known to be expressed mostly in the exocrine cells, including the salivary gland acinar cells. A naturally occurring point mutation (G308A, Gly103 > Asp103) was earlier found in the rat AQP5 gene [Murdiastuti, Purwanti, Karabasil, Li, Yao, Akamatsu, Kanamori and Hosoi (2006) Am. J. Physiol. 291, G1081-G1088]; in this mutant, the rate of initial saliva secretion under stimulated and unstimulated conditions is less than that for the wt (wild-type) animals. Here the mutant molecule was characterized in detail. Using the Xenopus oocyte system, we demonstrated the mutant AQP5 to have water permeability almost the same as that of the wt molecule. Mutant and wt AQP5s, tagged with GFP (green fluorescent protein; GFP-AQP5s) and expressed in polarized MDCK-II (Madin-Darby canine kidney II) cells, first appeared in the vesicular structure(s) in the cytoplasm, and were translocated to the upper plasma membrane or apical membrane during cultivation, with the mutant GFP-AQP5 being translocated less efficiently. Thapsigargin and H-89 both induced translocation in vitro of either molecule, whereas colchicine inhibited this activity; the fraction of cells showing apical localization of mutant GFP-AQP5 was less than that showing that of the wt molecule under any of the experimental conditions used. In the mutant SMG (submandibular gland) tissue, localization of AQP5 in the apical membrane of acinar cells was extremely reduced. Vesicular structures positive for AQP5 and present in the cytoplasm of the acinar cells were co-localized with LAMP2 (lysosome-associated membrane protein 2) or cathepsin D in the mutant gland, whereas such co-localizations were very rare in the wt gland, suggesting that the mutant molecules largely entered lysosomes for degradation. Replacement of highly conserved hydrophobic Gly103 with strongly hydrophilic Asp103 in rat AQP5, though it did not affect water permeability, may possibly have resulted in less efficient membrane trafficking and increased lysosomal degradation, leading to its lower expression in the apical membrane of the acinar cells in the SMG.
(キーワード)
Amino Acid Sequence / Animals / Aquaporin 5 / Cell Line / Cell Membrane / Dogs / Molecular Sequence Data / Mutation, Missense / Permeability / Point Mutation / Protein Transport / Rats / Sequence Alignment / Submandibular Gland / Water / Xenopus
Ahmad Azlina, Purevjav Javkhlan, Yuka Hiroshima, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : Roles of lysosomal proteolytic systems in AQP5 degradation in the submandibular gland of rats following chorda tympani parasympathetic denervation, American Journal of Physiology, Gastrointestinal and Liver Physiology, 299, 5, G1106-G1117, 2010.
(要約)
Chorda tympani denervation (CTD) of rats was earlier shown to result in loss of submandibular gland (SMG) weight (at only 1 wk) and in continued reduction in aquaporin 5 (AQP5) protein expression (until 4 wk), without affecting its mRNA synthesis (Li X, Azlina A, Karabasil MR, Purwanti N, Hasegawa T, Yao C, Akamatsu T, Hosoi K. Am J Physiol Gastrointest Liver Physiol 295: G112-G123, 2008). The present study indicated that despite elevation of bax, a proapoptosis protein, by CTD, the operation also increased the level of bcl-2, an antiapoptosis protein, in the SMG. Terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL assay) showed no increase in the number of apoptotic cells in the SMG. CTD, however, induced strongly and transiently (at 1-3 days) the protein expression of LC3B-II, a marker protein of autophagosomes, suggesting that the reduction in the gland weight was due to onset of autophagy by CTD. Upon CTD, Lamp2, a lysosomal marker, gradually increased in amount, reaching a peak at the 14th day. Immunohistochemical analysis revealed an increase in the number of lysosome-like structures positive for both AQP5 and Lamp2 in the acinar cells of the SMG after CTD; similar changes were observed also for AQP5 and LC3Bs. These data suggest that AQP5 in the SMG entered autophagosomes and/or lysosomes for degradation upon CTD. In vitro AQP5-degrading activity was found in the SMG extracts, and such activity was shown to be increased by CTD. Inhibitor experiments implied cathepsins B and L to be candidate enzymes for this degradation under normal and CTD conditions, respectively.
Chenjuan Yao, Nunuk Purwanti, Mileva Ratko Karabasil, Ahmad Azlina, Javkhlan Purevjav, Takahiro Hasegawa, Tetsuya Akamatsu, Toru Hosoi, Koichiro Ozawa and Kazuo Hosoi : Potential down-regulation of salivary gland AQP5 by LPS via cross-coupling of NF-κB and p-c-Jun/c-Fos, The American Journal of Pathology, 177, 2, 724-734, 2010.
(要約)
The mRNA and protein levels of aquaporin (AQP)5 in the parotid gland were found to be potentially decreased by lipopolysaccharide (LPS) in vivo in C3H/HeN mice, but only weakly in C3H/HeJ, a TLR4 mutant mouse strain. In the LPS-injected mice, pilocarpine-stimulated saliva production was reduced by more than 50%. In a tissue culture system, the LPS-induced decrease in the AQP5 mRNA level was blocked completely by pyrrolidine dithiocarbamate, MG132, tyrphostin AG126, SP600125, and partially by SB203580, which are inhibitors for IkappaB kinase, 26S proteasome, ERK1/2, JNK, and p38 MAPK, respectively. In contrast, the expression of AQP1 mRNA was down-regulated by LPS and such down-regulation was blocked only by SP600125. The transcription factors NF-kappaB (p65 subunit), p-c-Jun, and c-Fos were increased by LPS given in vivo, whereas the protein-binding activities of the parotid gland extract toward the sequences for NF-kappaB but not AP-1-responsive elements present at the promoter region of the AQP5 gene were increased by LPS injection. Co-immunoprecipitation by using antibody columns suggested the physical association of the three transcription factors. These results suggest that LPS-induced potential down-regulation of expression of AQP5 mRNA in the parotid gland is mediated via a complex(es) of these two classes of transcription factors, NF-kappaB and p-c-Jun/c-Fos.
Mileva Ratko Karabasil, Takahiro Hasegawa, Ahmad Azlina, Nunuk Purwanti, Javkhlan Purevjav, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : Trafficking of GFP-AQP5 chimeric proteins conferred with unphosphorylated amino acids at their PKA-target motif (152SRRTS) in MDCK-II cells, The Journal of Medical Investigation : JMI, 56, 1, 2, 55-63, 2009.
(要約)
Three constructs having mutated PKA-target motif at (152)SRRTS of AQP5, an exocrine type water channel, were prepared and fused to C-terminus of green fluorescence protein cDNA to examine the effects of blocking of phosphorylation at (152)SRRTS (a consensus PKA-target motif of AQP5) on translocation or trafficking of the chimeric proteins expressed in the Madin-Darby canine kidney-II (MDCK-II) cells. H-89 treatment increased translocation of wild-type GFP-AQP5 to the apical membrane. All 3 mutant molecules translocated 1.5 to 2 times more than the control wild-type GFP-AQP5. Colchicine but not cytochalasin B inhibited the translocation of wild-type GFP-AQP5. Present results suggest dephosphorylation of this consensus sequence increase GFP-AQP5 translocation, and that microtubules but not microfilaments are involved in this event.
(キーワード)
Amino Acid Motifs / Amino Acids / Animals / Aquaporin 5 / Cell Line / Cell Membrane / Chimera / Colchicine / Cyclic AMP-Dependent Protein Kinases / Cytochalasin B / Dogs / Green Fluorescent Proteins / Isoquinolines / Kidney / Phosphorylation / Protein Transport / Sulfonamides
Shingo Kurabuchi and Kazuo Hosoi : Immunocytochemical study of granular duct cells with a hormonally enhanced granular cell phenotype in the mouse parotid gland, Odontology, 97, 1, 57-61, 2009.
(要約)
In the parotid glands (PGs) of intact male mice (12 weeks of age, ICR strain), immunofluorescence labels for a true tissue kallikrein, mK1, and for nerve growth factor (NGF) were recognized through the subluminal edges of the striated duct (SD) segments and interlobular duct segments. Because of their small size, secretory granules were not detectable by light microscopy in any of the duct cells. Full-fledged granular cells, containing large secretory granules that were visible by light microscopy, were induced in the SD segments of male mice after the injection of 5alpha-dehydrotestosterone (DHT) and triiodothyronine (T(3)), given either alone or in combination every other day for 2 weeks. A stronger effect was detected in the mice that were concomitantly injected with DHT and T(3), and more abundant, fully developed granular cells appeared in the SD segments of these mice. These full-fledged granular cells were immunoreactive for mK1, NGF, and epidermal growth factor, but not for renin. The present results indicate that some of the SD cells with small granules in the mouse PG can develop a granular cell phenotype, producing more kinds of growth factors, as a result of the actions of androgen and thyroid hormone.
Tetsuya Akamatsu, Ahmad Azlina, Nunuk Purwanti, Mileva Ratko Karabasil, Takahiro Hasegawa, Chenjuan Yao and Kazuo Hosoi : Inhibition and transcriptional silencing of a subtilisin-like proprotein convertase, PACE4/SPC4, reduces the branching morphogenesis of and AQP5 expression in rat embryonic submandibular gland, Developmental Biology, 325, 2, 434-443, 2009.
(要約)
The submandibular gland (SMG) develops through the epithelial-mesenchymal interaction mediated by many growth/differentiation factors including activin and BMPs, which are synthesized as inactive precursors and activated by subtilisin-like proprotein convertases (SPC) following cleavage at their R-X-K/R-R site. Here, we found that Dec-RVKR-CMK, a potent inhibitor of SPC, inhibited the branching morphogenesis of the rat embryonic SMG, and caused low expression of a water channel AQP5, in an organ culture system. Dec-RVKR-CMK also decreased the expression of PACE4, a SPC member, but not furin, another SPC member, suggesting the involvement of PACE4 in the SMG development. Heparin, which is known to translocate PACE4 in the extracellular matrix into the medium, and an antibody specific for the catalytic domain of PACE4, both reduced the branching morphogenesis and AQP5 expression in the SMG. The inhibitory effects of Dec-RVKR-CMK were partially rescued by the addition of recombinant BMP2, whose precursor is one of the candidate substrates for PACE4 in vivo. Further, the suppression of PACE4 expression by siRNAs resulted in decreased expression of AQP5 and inhibition of the branching morphogenesis in the present organ culture system. These observations suggest that PACE4 regulates the SMG development via the activation of some growth/differentiation factors.
Shingo Kurabuchi, Edward W Gresik, Chenjuan Yao and Kazuo Hosoi : Hypophysectomy and hormonal therapy modulate mK1-immunoreactive duct cells in the mice sublingual glands, Journal of Molecular Histology, 39, 5, 499-507, 2008.
(要約)
The immunocytochemical localization of a true tissue kallikrein, mK1, in mouse sublingual glands (SLGs) was examined following hypophysectomy and hormonal replacement therapy. In the glands of intact mice (14 weeks of age), mK1 was detected in the striated ducts (SDs). Full-fledged granular cells were scattered in the SDs of male mice (but not in those of female mice), showing a cellular mosaic distribution of mK1 with some being positive and others being negative. mK1 was also detected in transitional-type granular cells, though the secretory granules were too small and scarce to be visible by a light microscopy. Hypophysectomy in male mice resulted in the atrophy and loss of secretory granules in many SD cells. Granulation recovered after the repeated injection of 5alpha-dihydrotestosterone (DHT), 3,5,3'-triiodo-L: thyronine (T3), and dexamethasone (Dex), given either alone or in combination to the hypophysectomized mice. The concomitant injection of DHT and T3, with or without Dex, resulted in the reappearance of the full-fledged granular cells, only some of which were mK1-positive. Electron microscopy revealed mK1 to be present exclusively in the secretory granules of these mK1-positive cells, and no ultrastructural differences were observed between mK1-positive and mK1-negative full-fledged granular cells. These results show that the differentiation of the granular cell phenotype in the mouse SLG duct system requires the concomitant action of androgen and thyroid hormone and retards mK1 synthesis.
Xuefei Li, Ahmad Azlina, Mileva Ratko Karabasil, Nunuk Purwanti, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : Degradation of submandibular gland AQP5 by parasympathetic denervation of chorda tympani and its recovery by cevimeline, an M3 muscarinic agonist, American Journal of Physiology, Gastrointestinal and Liver Physiology, 295, 1, G112-G123, 2008.
(要約)
By chorda tympani denervation (CTD, parasympathectomy), the aquaporin 5 (AQP5), but not AQP1, protein level in the rat submandibular gland (SMG) was significantly decreased, dropping to 37% of that of the contralateral gland at 4 wk. The protein levels of AQP5 and AQP1 were not significantly affected by denervation of the cervical sympathetic trunk (sympathectomy). Administration of cevimeline hydrochloride, an M3 muscarinic receptor agonist (10 mg/kg for 7 days po), but not pilocarpine (0.3 mg/kg for 7 days po), recovered the AQP5 protein level reduced by CTD and increased the AQP1 protein level above the control one. The mRNA level of AQP5 was scarcely affected by CTD and cevimeline hydrochloride administration. Administration of chloroquine (50 mg/kg for 7 days po), a denaturant of lysosomes, increased the AQP5 protein level reduced by CTD. An extract obtained from the submandibular lysosomal fraction degraded the AQP5 protein in the total membrane fraction in vitro. These results suggest the possible regulation of the AQP5 protein level in the SMG by the parasympathetic nerves/M3 muscarinic receptor agonist and imply the involvement of lysosomal enzymes, but not a transcriptional mechanism, in this regulation.
Tetsuya Akamatsu, Nunuk Purwanti, Mileva Ratko Karabasil, Xuefei Li, Chenjuan Yao, Norio Kanamori and Kazuo Hosoi : Temporospatially regulated expression of subtilisin-like proprotein convertase PACE4 (SPC4) during development of the rat submandibular gland, Developmental Dynamics, 236, 1, 314-320, 2007.
(要約)
The temporospatial expression of PACE4, a member of the mammalian subtilisin-like proprotein convertase family involved in the activation of growth/differentiation factors, was investigated by in situ hybridization during the development of the rat submandibular gland (SMG). At the initiation stage (day 15.5 of gestation; E15), PACE4 was intensely expressed in the submandibular epithelium, but weakly expressed in the mesenchymal cells. At E16 when the branching morphogenesis becomes obvious, the expression of PACE4 in the mesenchyme was further decreased, although its level in the submandibular epithelium had not changed remarkably from that at E15. During the next stage of embryonic development (E17-E20), PACE4 was expressed in the cells derived from the submandibular epithelium, which include the proacinar, terminal tubular, and presumptive ductal cells. In the perinatal SMG, PACE4 was still expressed intensely in the terminal portion of the SMG containing the proacinar and terminal tubular cells, whereas its expression in the ductal cells was obviously decreased at the second postnatal day (P2) and at P6. Acinar cells expressing no PACE4 appeared, and their numbers increased following their development (P9-P20). At P30 when the PACE4 expression in the acinar cells was completely suppressed, its expression in the ductal cells became intense again. This temporospatially regulated expression of PACE4 suggests its apparent association with the proliferation, differentiation, and establishment of functional acinar and ductal cells of the SMG.
Kwartarini Murdiastuti, Nunuk Purwanti, Mileva Ratko Karabasil, Xuefei Li, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : A naturally occurring point mutation in the rat aquaporin 5 gene, influencing its protein production by and secretion of water from salivary glands, American Journal of Physiology, Gastrointestinal and Liver Physiology, 291, 6, GI1081-GI1088, 2006.
(要約)
A greater than twofold diversity in the expression level of aquaporin 5 (AQP5) has been observed in the membrane fraction of the submandibular gland (SMG) in Sprague-Dawley rats (Murdiastuti K, Miki O, Yao C, Parvin MN, Kosugi-Tanaka C, Akamatsu T, Kanamori N, and Hosoi K. Pflügers Arch 445: 405-412, 2002). In the present study, breeding between brother and sister rats was repeated within high AQP5 producers and low ones to obtain inbred offspring. High- and low-producer rats from 3rd to 18th generations were used for experiments. By Western blotting, levels of AQP5 proteins in the parotid and lacrimal glands, and lungs were all low in low producers, whereas they were all high in high producers, implying genetic variations of the gene for this water channel. Despite this implication, AQP5 mRNA levels were almost the same between the two groups by Northern blotting, suggesting the irrelevance of transcriptional regulation for this diversity. AQP5 cDNAs from the SMGs of the two groups were sequenced. The nucleotide sequence of AQP5 cDNA from low producers indicated the existence of a point mutation at nt 308 (G308A), leading to a replacement of (103)Gly with (103)Asp in the third transmembrane domain, but no alteration was detected in the Kozak area. The existence of such a mutation was confirmed by the assessment of genomic DNA also. This mutation may have resulted in an abnormal membrane insertion or ineffective trafficking of AQP5, since the rats having this mutation showed extremely low membrane expression of AQP5 in the SMG acinar cells and decreased water secretion from their salivary glands.
(キーワード)
Amino Acid Sequence / Animals / Aquaporin 5 / Base Sequence / Body Water / Molecular Sequence Data / Point Mutation / Rats / Salivary Glands / Sequence Homology, Amino Acid / Structure-Activity Relationship
Chisato Kosugi, Xuefei Li, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Protein kinase A-regulated membrane trafficking of a green fluorescent protein-aquaporin 5 chimera in MDCK cells, Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1763, 4, 337-344, 2006.
(要約)
The green fluorescent protein (GFP) of the jellyfish, Aeqorea victoria, was used as an autofluorescent tag to track the trafficking of aquaporin 5 (AQP5), an exocrine gland-type water channel. Two groups of chimeric proteins were constructed; one in which GFP was fused to the amino-terminus of AQP5 (GFP-AQP5) and the other, in which it was fused to the carboxyl terminus of it (AQP5-GFP). In each group, 2 chimeras were produced, a wild-type AQP5 with its normal sequence and a mutant AQP5 having a mutated amino acid at 259, i.e., GFP-AQP5-T259A and AQP5-GFP-T259A. They were used to transfect Madin-Darby canine kidney (MDCK) cells. The GFP-AQP5 chimera was localized in the intracellular vesicles, which trafficked to the plasma membrane in response to N(6), 2'-O-dibutyryladenosine 3', 5'-cyclic monophosphate (dbcAMP). Membrane trafficking was inhibited by N-[2-(p-bromocinnamylamino)ethyl]-5-isoquimolinesulfonamide (H-89) but not by palmitoyl-dl-carnitine chloride (PCC). In contrast, the AQP5-GFP chimera expressed in MDCK cells was localized constitutively on the plasma membrane. The cellular localization of the latter chimera was not affected by stimulation with dbcAMP in the presence or absence of H-89 or PCC. Replacement of Thr-259 with Ala-259 did not affect the dbcAMP-induced translocation of the chimeric protein, suggesting that phosphorylation of Thr-259 was not necessary for AQP5 trafficking under the present experimental conditions. Thus, the GFP-AQP5 chimera will be a useful tool to study AQP5 trafficking in vitro, whereas the constitutive membrane localization of the AQP5-GFP chimera suggests the importance of the carboxyl terminus of the AQP5 protein for its sorting, whether it is translocated to intracellular vesicles or to the plasma membrane.
(キーワード)
Animals / Aquaporin 5 / Cell Line / Cell Membrane / Cyclic AMP-Dependent Protein Kinases / Dogs / Green Fluorescent Proteins / Protein Transport / Recombinant Fusion Proteins
Masataka Murakami, Kwartarini Murdiastuti, Kazuo Hosoi and Adrian E Hill : AQP and the control of fluid transport in a salivary gland, The Journal of Membrane Biology, 210, 2, 91-103, 2006.
(要約)
Experiments were performed with the perfused rat submandibular gland in vitro to investigate the nature of the coupling between transported salt and water by varying the osmolarity of the source bath and observing the changes in secretory volume flow. Glands were submitted to hypertonic step changes by changing the saline perfusate to one containing different levels of sucrose. The flow rate responded by falling to a lower value, establishing a new steady-state flow. The rate changes did not correspond to those expected from a system in which fluid production is due to simple osmotic equilibration, but were much larger. The changes were fitted to a model in which fluid production is largely paracellular, the rate of which is controlled by an osmosensor system in the basal membrane. The same experiments were done with glands from rats that had been bred to have very low levels of AQP5 (the principal aquaporin of the salivary acinar cell) in which little AQP5 is expressed at the basal membrane. In these rats, salivary secretion rates after hypertonic challenges were small and best modelled by simple osmotic equilibration. In rats which had intermediate AQP5 levels the changes in flow rate were similar to those of normal rats although their AQP5 levels were reduced.Finally, perfused normal glands were subject to retrograde ductal injection of salines containing different levels of Hg(2+) ions (0, 10 and 100 microM: ) which would act as inhibitors of AQP5 at the apical acinar membrane. The overall flow rates were progressively diminished with rising Hg(2+) concentration, but after hypertonic challenge the changes in flow rates were unchanged and similar to those of normal rats. All these results are difficult to explain by a cellular osmotic model but can be explained by a model in which paracellular flow is controlled by an osmosensor (presumably AQP5) present on the basal membrane.
Chenjuan Yao, Mileva Ratko Karabasil, Nunuk Purwanti, Xuefei Li, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Tissue kallikrein mK13 is a candidate processing enzyme for the precursor of interleukin-1b in the submandibular gland of mice, The Journal of Biological Chemistry, 281, 12, 7968-7976, 2006.
(要約)
By using Western blot analysis, high levels of 17.5- and 20-kDa interleukin-1beta (IL-1beta) proteins were detected in the submandibular gland (SMG) of mice. Despite this fact, the amount of pro-IL-1beta protein, a precursor of IL-1beta, with a molecular size of 35 kDa in this tissue was below the detectable level, although strong expression of pro-IL-1beta mRNA was observed. A large amount of 17.5-kDa IL-1beta also appeared in the saliva of mice injected with lipopolysaccharide, suggesting that this IL-1beta is a secretory form produced by the SMG. The protein for IL-1beta-converting enzyme, a processing enzyme for pro-IL-1beta, was expressed only at a low level in the SMG as compared with its level in various epithelial tissues or lipopolysaccharide-stimulated macrophages. On the other hand, mK1, mK9, mK13, and mK22, members of the kallikrein family, were detected strongly in the SMG but not in other tissues. By incubation with mK13, but not with mK1, mK9, or mK22, the 35-kDa pro-IL-1beta was cleaved into two major products with molecular masses of 17.5 and 22 kDa, and production was inhibited by phenylmethylsulfonyl fluoride, a serine protease inhibitor, but not by IL-1beta-converting enzyme inhibitors. A peptide segment corresponding to amino acid residues 107-121 of mouse pro-IL-1beta (107WDDDDNLLVCDVPIR) was cleaved by incubation with mK13, generating two peptides, 107WDDDDNL and 114LVCDVPIR. Therefore, kallikrein mK13 would appear to hydrolyze pro-IL-1beta between its Leu113 and Leu114 residues. The results of immunohistochemistry and an autonomic therapy experiment showed that IL-1beta and kallikrein mK13 were co-localized in the secretory granules of granular convoluted tubular cells. Our present results thus suggest kallikrein mK13 is a plausible candidate for the processing enzyme for pro-IL-1beta in the SMG of mice.
Mohammad Alizadeh, Fusao Ota, Kazuo Hosoi, Makoto Kato, Tohru Sakai and Mohammed A. Satter : Altered allergic cytokine and antibody response in mice treated with Bisphenol A, The Journal of Medical Investigation : JMI, 53, 1, 2, 70-80, 2006.
(要約)
The objective of this study was to elucidate if Bisphenol A (BPA) administration modulates T helper (Th) cell component of immune responses in a mouse challenged with ovalbumin (OVA), a major food antigen. BALB/c mice, (6 weeks old, female) were orally given either OVA (OVA-fed) or water (Water-fed), immunized intraperitoneally with OVA and injected with either BPA in corn oil or the vehicle alone. After subsequent 2nd immunization, serum titers of total IgE, OVA-specific IgE, IgG, IgG1 IgG2a and ability of their splenocytes for production of interferon (IFN) -gamma, interleukin (IL) -4 and IL-12 were examined by ELISA. Lymphocyte proliferation assay against concanavalin A (Con A) or OVA was also performed for 3H-Thymidine incorporation. In Water-fed groups, treatment with BPA resulted in lower titers of total IgE (P<0.01) and higher levels IgG2a (P<0.05) followed by a higher IFN-gamma (P<0.05) and IL-12 (P<0.05) with an intact IL-4. When OVA-fed groups were examined, the compound did not change production of total and OVA-specific IgE and -IgG2a but resulted in lower production of IFN-gamma (P<0.05). Also, BPA resulted in impaired lymphocyte proliferation to Con A in Water-fed groups (P<0.05) but not in tolerated animals. The findings indicate that BPA results in augmentation of Th1 immune responses but no significant effect on an established tolerance to OVA.
Keiko Tsumura, Xuefei Li, Kwartarini Murdiastuti, Most Nahid Parvin, Tetsuya Akamatsu, Chenjuan Yao, Norio Kanamori, Kiyotoshi Inenaga, Hiroshi Yamashita and Kazuo Hosoi : Downregulation of AQP2 expression in the kidney of polydipsic STR/N mice, American Journal of Physiology, Renal Physiology, 290, 2, F478-F485, 2006.
(要約)
Aquaporin-2 (AQP2) is responsible for the concentration of urine in the kidney collecting tubule under the regulation of vasopressin. The mRNA level of this water channel in polydipsic STR/N mice was extremely reduced compared with that in normal ICR mice. In male mice, reduction of the AQP2 mRNA level was not evident at 3 wk of age, at which time water intake was not increased. At 10 wk of age, however, the AQP2 mRNA level was reduced to 10% of that in control mice, whereas water intake was increased by 36%. At 44 wk, the water intake became five times that of the control ICR mice, and the AQP2 mRNA level in these polydipsic mice was only approximately 5% of control. Similar changes were observed in the AQP2 protein level, suggesting that the mRNA level of AQP2 reflects the protein level of AQP2. These inverse changes in the AQP2 mRNA level and water intake were also evident in female mice. The data imply that polydipsia in STR/N mice may have affected AQP2 mRNA transcription in the kidney, resulting in reduced AQP2 expression, which would contribute to a reduction in overretention of water.
Chenjuan Yao, Xuefei Li, Murdiastuti Kwartarini, Chisato Kosugi, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Lipopolysaccharide-induced elevation and secretion of Interleukin-1b in the submandibular gland of male mice, Immunology, 116, 2, 213-222, 2005.
(要約)
The intraperitoneal injection of lipopolysaccharide (LPS) (400 microg/kg body weight) induced the expression of mRNAs of inflammatory cytokines such as interleukin (IL)-1beta, IL-6 and tumour necrosis factor (TNF)-alpha in the submandibular gland (SMG) of C3H/HeN mice but not that of C3H/HeJ mice, a mutant strain for Toll-like receptor-4 (TLR-4(-) mutant). The mRNA levels of these cytokines in the SMG of the wild-type mice increased as early as 3 hr after injection, peaked at 3-6 hr, and had decreased again by 24 hr. In this study, we particularly focused on IL-1beta, and induction by this endotoxin was investigated in detail. Denervation of the superior cervical trunk and chorda tympani nerve did not diminish the LPS-induced elevation of IL-1beta mRNA in the SMG, indicating the irrelevance of the central nervous system in this induction. TLR-4 mRNA and protein were shown to be strongly expressed in the SMG, suggesting the direct action of LPS on this gland. IL-1beta proteins were localized in the secretory granules of granular convoluted tubular (GCT) cells, and their molecular weights in the gland were 17.5 and 20 kDa. IL-1beta of the same size appeared in the saliva 6 hr after LPS injection in C3H/HeN but not in C3H/HeJ mice. The present study thus suggests that IL-1beta, an inflammation cytokine, is induced and secreted into the saliva in response to endotoxin injected intraperitoneally.
Chenjuan Yao, Wei Wei, Xuefei Li and Kazuo Hosoi : Acute phase protein induction by experimental inflammation in the salivary gland, Journal of Oral Pathology & Medicine, 34, 6, 364-367, 2005.
(要約)
The submandibular gland (SMG) is a major salivary gland, which plays an important role in maintenance the oral health. In this study, we intended to explore the role of the SMG's defense system of the animals in which experimental inflammation is induced. The levels of mRNAs for inflammation cytokines and acute phase proteins were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). The mRNAs for acute phase proteins were found to be increased in the SMG and extraorbital and intraorbital lacrimal gland (ELG and ILG) of rats at 24 h after subcutaneous injection of turpentine oil. The induction of mRNA for these inflammatory proteins by turpentine oil was preceded by a transient increase in the level of mRNAs for IL-1beta, IL-6 and TNF-alpha at 6 h after subcutaneous injection of the oil. Such cytokine induction was similarly seen by lipopolysaccharide (LPS) injection, and involvement of Toll-like receptor 4 (TLR4) was strongly suggested from experiment using C3H/HeJ mice, a TLR4-deficient mutant strain. The up-regulation of acute phase proteins and inflammation cytokines in the SMG, ELG and ILG by experimental inflammation suggests the existence of a strict defense system via the innate immune system in the SMG and other exocrine gland.
Most Nahid Parvin, Shingo Kurabuchi, Kwartarini Murdiastuti, Chenjuan Yao, Chisato Kosugi, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Subcellular redistribution of AQP5 by vasoactive intestinal polypeptide (VIP) in the Brunner's gland of the rat duodenum, American Journal of Physiology, Gastrointestinal and Liver Physiology, 288, 6, G1283-G1291, 2005.
(要約)
Aquaporin (AQP)5, an exocrine-type water channel, was detected in the rat duodenum by Western blot analysis, and was localized by immunohistochemistry in the secretory granule membranes as well as in the apical and lateral aspects of the plasma membrane of Brunner's gland cells. Incubation of duodenal slices with vasoactive intestinal polypeptide (VIP) in vitro significantly increased the amount of AQP5 in the apical membrane fraction in a dose- and time-dependent manner with the amount reaching a plateau at 100 nM VIP and becoming near maximal after a 30-s incubation. Protein kinase inhibitors, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine dihydrochloride (H-7, 50 muM), and N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H-89; PKA-specific, 1 muM) blocked this increase, but PKC-specific inhibitor calphostin C did not, implying the involvement of PKA but not PKC in this cellular event. Intravenous injection with VIP (40 mug/kg body wt) provoked dilation of the lumen of the Brunner's gland at 2 and 7 min and increased the staining intensity of AQP5 in the apical and lateral membranes. AQP1 (both nonglycosylated and glycosylated forms) was also found to localize in the apical and basolateral membranes of cells of Brunner's gland. VIP, however, did not provoke any significant change in the AQP1 level in the apical membrane, as judged from the results of the above in vitro and in vivo experiments. These results suggest that VIP induced the exocytosis of granule contents and simultaneously caused translocation of AQP5 but not of AQP1 to the apical membrane in Brunner's gland cells.
Shingo Kurabuchi, Edward W Gresik and Kazuo Hosoi : Additive and/or synergistic action (downregulation) of androgens and thyroid hormones on the cellular distribution and Localization of a true tissue kallikrein, mK1, in the mouse submandibular gland, The Journal of Histochemistry and Cytochemistry, 52, 11, 1437-1446, 2004.
(要約)
We investigated the effects of 5alpha-dihydrotestosterone (DHT), 3,5,3'-triiodo-l-thyronine (T(3)), and dexamethasone (Dex) on the expression of mK1 in the granular convoluted tubule (GCT) cells of the submandibular gland (SMG) of hypophysectomized (Hypox) male mice by indirect enzyme-labeled antibody and immunogold antibody methods for light and electron microscopy. Hypox resulted in considerable atrophy of the GCT cells, which were always immunoreactive for mK1, and the cells were characterized by apical small dense secretory granules labeled with gold particles suggesting the presence of mK1, small Golgi apparatus, sparse rough endoplasmic reticulum (RER), and developed basal infoldings. Each of the hormones, DHT, T(3), and Dex, enhanced the GCT phenotype to various degrees in Hypox male mice. Both DHT alone and T(3) alone moderately inhibited mK1 synthesis by increasing the number of mK1-immunonegative GCT cells in Hypox males, but Dex alone had no inhibitory effect on mK1 synthesis. A significant trophic effect on GCT cells was induced by combined injection of DHT and T(3) or of all three hormones, and was reflected in the appearance of abundant large secretory granules, well-developed Golgi apparatus and RER, and reduced basal infoldings. Only a few such GCT cells were immunopositive for mK1, and the pattern of immunopositive and immunonegative cells very closely resembled the mosaic pattern seen in normal male GCTs. These findings suggested that the sexual dimorphism of mK1 expression and the morphological appearance of GCT cells can be induced by treatment with two hormones, DHT and T(3), but not by either of them alone. T(3) appears to have a permissive effect on committed GCT cells that results in downregulation of mK1 expression in these cells.
Shingo Kurabuchi and Kazuo Hosoi : Immunocytochemical localization of mK1, a true tissue kallikrein, in the mouse parotid gland: sexual dimorphism and effects of castration and hypophysectomy, Odontology, 92, 1, 73-76, 2004.
(要約)
In the normal parotid glands of mice at 12 weeks of age, mK1, a true tissue kallikrein, was detected at the apical rim of the striated ducts (SDs). Sexual dimorphism in the immunostaining intensity in parotid glands was seen, i.e., immunostaining was more intense in males than in females. Under electron microscopy, secretory granules, being small in size, and condensed at the subluminal cytoplasm, were labeled with immunogold particles showing the presence of mK1. These secretory granules were rather abundant and large in males. Castration in males reduced the immunoreactivity of mK1 in the SD cells because of a decrease in the number and size of secretory granules as revealed by electron microscopy. Hypophysectomy in male mice resulted in considerable loss of immunoreactivity for mK1, which was characterized under electron microscopy by complete disappearance or significant reduction of secretory granules in many SD cells. These results suggest that mK1 expression in the SD cells of murine parotid glands is regulated by pituitary-dependent hormones, and sexual dimorphism of mK1 expression is regulated by androgens.
Sandro E. Fogaça, Robson L. Melo, Daniel C. Pimenta, Kazuo Hosoi, Luiz Juliano and Maria A. Juliano : Difference in substrate and inhibitor sequence specificity of human, mouse and rat tissue kallikreins, The Biochemical Journal, 380, Pt3, 775-781, 2004.
(要約)
The kininogenase activities of mouse (mK1), rat (rK1) and human (hK1) tissue kallikreins were assayed with the bradykinin-containing synthetic peptides Abz-MTEMARRPPGFSPFRSVTVQNH2 (where Abz stands for o-aminobenzoyl) and Abz-MTSVIRRPPGFSPFRAPRV-NH2, which correspond to fragments Met374-Gln393 and Met375-Val393 of mouse and rat LMWKs (low-molecular-mass kininogens) with the addition of Abz. Bradykinin was released from these peptides by the mK1- and rK1-mediated hydrolysis of Arg-Arg and Arg-Ser (or Arg-Ala) peptide bonds. However, owing to preferential hydrolysis of Phe-Arg compared with the Arg-Ala bond in the peptide derived from rat LMWK, hK1 released bradykinin only from the mouse LMWK fragment and preferentially released des-[Arg9]bradykinin from the rat LMWK fragment (Abz-MTSVIRRPPGFSPFRAPRV-NH2). The formation of these hydrolysis products was examined in more detail by determining the kinetic parameters for the hydrolysis of synthetic, internally quenched fluorescent peptides containing six N- or C-terminal amino acids of bradykinin added to the five downstream or upstream residues of mouse and rat kininogens respectively. One of these peptides, Abz-GFSPFRAPRVQ-EDDnp (where EDDnp stands for ethylenediamine 2,4-dinitrophenyl), was preferentially hydrolysed at the Phe-Arg bond, confirming the potential des-[Arg9]bradykinin-releasing activity of hK1 on rat kininogen. The proline residue that is two residues upstream of bradykinin in rat kininogen is, in part, responsible for this pattern of hydrolysis, since the peptide Abz-GFSPFRASRVQ-EDDnp was preferentially cleaved at the Arg-Ala bond by hK1. Since this peptidase accepts the arginine or phenylalanine residue at its S1 subsite, this preference seems to be determined by the prime site of the substrates. These findings also suggested that the effects observed in rats overexpressing hK1 should consider the activation of B1 receptors by des-[Arg9]bradykinin. For further comparison, two short internally quenched fluorescent peptides that bind to hK1 with affinity in the nM range and some inhibitors described previously for hK1 were also assayed with mK1 and rK1.
Tetsuya Akamatsu, Most Nahid Parvin, Kwartarini Murdiastuti, Chisato Kosugi, Chenjuan Yao, Osamu Miki, Norio Kanamori and Kazuo Hosoi : Expression and localization of aquaporins, members of the water channel family, during development of the rat submandibular gland., Pflügers Archiv : European Journal of Physiology, 446, 6, 641-651, 2003.
Kwartarini Murdiastuti, Osamu Miki, Chenjuan Yao, Most Nahid Parvin, Chisato Kosugi, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Divergent expression and localization of aquaporin 5, an exocrine-type water channel, in the submandibular gland of Sprague-Dawley rats., Pflügers Archiv : European Journal of Physiology, 445, 3, 405-412, 2002.
Shingo Kurabuchi, Kazuo Hosoi and Edward W Gresik : Developmental and androgenic regulation of the immunocytochemical distribution of mK1, a true tissue kallikrein, in the granular convoluted tubule of the mouse submandibular gland, The Journal of Histochemistry and Cytochemistry, 50, 2, 135-145, 2002.
(要約)
The action of androgens on the immunocytochemical distribution of mK1, a true tissue kallikrein, was examined in the submandibular gland (SMG) of developing and adult mice by indirect enzyme-labeled and immunogold-labeled antibody methods for light and electron microscopy, respectively. In both sexes at 3 weeks of age, essentially all of the immature granular convoluted tubule (GCT) cells were uniformly immunostained. At 4 weeks of age (the onset of puberty), morphological differences between the two sexes appeared in the GCTs, in which some cells became immunonegative. Thereafter, the immunonegative GCT cells became more abundant in the SMG of males than of females and considerable intercellular variation in staining intensity for mK1 was seen, especially in males. A few slender GCT cells with strong immunoreactivity appeared in GCT segments only in males. Castration of males resulted in an increase in the number of immunopositive GCT cells, whereas administration of dihydrotestosterone (DHT) decreased the number of immunopositive GCT cells in the SMGs of both sexes. Slender GCT cells immunoreactive for mK1 were seen in females treated with DHT for 6 days. However, there were no immunostained slender GCT cells in female SMGs after injection of DHT for 2 weeks. Immunoelectron microscopy disclosed this type of cell in male SMGs, which closely resembles immature GCT cells of prepubertal mice, with a few small secretory granules uniformly labeled with gold particles, a sparse Golgi apparatus and RER, and basal infoldings. In mature male SMGs and in SMGs of DHT-treated females and castrated males, typical GCT cells had a well-developed Golgi apparatus and a net-like RER but few to no basal infoldings, whereas in the female gland equivalent cells had moderately developed RER and some basal infoldings. These results suggest that mK1 is one of the enzymes characteristically present in immature GCT cells and that its synthesis is inhibited in part by androgens, resulting in decreased numbers of immunopositive cells.
Most Nahid Parvin, Keiko Tsumura, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Expression and localization of AOP5 in the stomach and duodenum of the rat., Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1542, 1-3, 116-124, 2002.
Norio Kanamori, Tetsuya Akamatsu, Chisato Kosugi, Most Nahid Parvin and Kazuo Hosoi : Methylene blue as a vascular filling dye., ITE Leters on Batteries, New Technologies & Medicine (with News), 3, 1, 75-77, 2002.
Shingo Kurabuchi, Kazuo Hosoi and Edward W Gresik : Androgen regulation of the cellular distribution of the true tissue kallikrein mK1 in the submandibular gland of the mouse, The Journal of Histochemistry and Cytochemistry, 49, 6, 801-802, 2001.
(要約)
The kallikrein gene family encodes for at least four different proteases in the mouse submandibular gland (SMG): mK1 (true tissue kallikrein), mK9, mK13, and mK22. These enzymes and many other biologically active proteins are synthesized by the granular convoluted tubule (GCT), a specialized segment of the SMG duct system. The GCT is under multihormonal regulation by androgens, thyroid hormones, and adrenocortical hormones. Androgens suppress synthesis of mK1 in the SMG but enhance expression of the other three kallikreins. We prepared an antibody with limited immunoreactivity for mK1 and used it to examine the effects of androgen status on the distribution of this isozyme in the SMGs of developing and mature mice by immunoperoxidase staining for the light microscope and immunogold labeling for the electron microscope. In prepubertal mice, every immature GCT cell contains mK1, confined to an accumulation of small granules in the subluminal cytoplasm. In mature mice, not every GCT cell contains mK1, and in those cells that do there is considerable intergranular variation in the intensity of staining for mK1. GCT cells containing mK1 are much more abundant in the glands of females than of males, resulting in a peculiar sexually dimorphic mosaic distribution of this isozyme in the mature SMG. Castration of adult males increases the number of GCT cells expressing mK1. Administration of androgen to intact or castrated males or to intact females reduces the number of cells staining for mK1. In all cases, immunogold labeling for mK1 is confined to secretory granules. No fine structural differences were noted between cells that were positively or negatively stained for mK1. Therefore, although GCT cells appear to be composed of a uniform population of cells on the basis of morphology alone, they are not homogeneous in their content of secretory proteins. These results indicate that androgen regulation of GCT cells is more complex than has been appreciated to date.
(キーワード)
Androgens / Animals / Antibody Specificity / Female / Immunoenzyme Techniques / Isoenzymes / Male / Mice / Phenotype / Sex Characteristics / Sexual Maturation / Submandibular Gland / Tissue Distribution / Tissue Kallikreins
Wei Wei, Most Nahid Parvin, Keiko Tsumura, Tetsuya Akamatsu, Jun Tada, Norio Kanamori and Kazuo Hosoi : Induction of C-reactive protein, serum amyloid P component, and kininogens in the submandibular and lacrimal glands of rats with experimentally induced inflammation., Life Sciences, 69, 3, 359-368, 2001.
Most Nahid Parvin, Kwartarini Murdiastuti, Tetsuya Akamatsu, Norio Kanamori, Norihiko Maeda and Kazuo Hosoi : Expression and locarization of AQP4 in the crypt epithelial cells of the rat duodenum., Biomedical Research (India), 12, 2, 77-82, 2001.
Norio Kanamori, Tetsuya Akamatsu, Jun Tada, Keiko Tsumura, Most Nahid Parvin, Kwartarini Murdiastui, Wei Wei and Kazuo Hosoi : Search for the dye that stains the vascular system., ITE Leters on Batteries, New Technologies & Medicine (with News), 2, 1, 116-119, 2001.
Tetsuya Akamatsu, Yoshiko Matsuda, Keiko Tsumura, Jun Tada, Most Nahid Parvin, Wei Wei, Norio Kanamori and Kazuo Hosoi : Highly regulated expression of subtilisin-like proprotein convertase PACE4 (SPC4) during dentinogenesis., Biochemical and Biophysical Research Communications, 272, 2, 410-415, 2000.
Kazuo Hosoi, Sachiko Matsuura, Keiko Tsumura, Wei Wei, Most Nahid Parvin, Jun Tada, Tetsuya Akamatsu, Norio Kanamori and Kazuo Suzuki : Expression of kininogens in the connective tissue-type mast cells of the rats., Immunology, 101, 4, 531-540, 2000.
Jun Tada, Takamasa Sawa, Naoki Yamanaka, Masayuki Shono, Tetsuya Akamatsu, Keiko Tsumura, Most Nahid Parvin, Norio Kanamori and Kazuo Hosoi : Involvement of vesicle-cytoskeleton interaction in AQP5 trafficking in AQP5-gene-transfected HSG cells., Biochemical and Biophysical Research Communications, 266, 2, 443-447, 1999.
Tetsuya Akamatsu, Yoshiko Matsuda, Keiko Tsumura, Jun Tada, Most Nahid Parvin, Norio Kanamori and Kazuo Hosoi : Subtilisin-like proprotein convertase PACE4 (SPC4) is a candidate processing enzyme of bone morphogenetic proteins during tooth formation, Developmental Dynamics, 216, 4-5, 481-488, 1999.
Shingo Kurabuchi, Jun Tada, Edward W Gresik and Kazuo Hosoi : An unusual sexually dimorphic mosaic distribution of a subset of kallikreins in the granular convoluted tubule of the mouse submandibular gland detected by an antibody with restricted immunoreactivity, The Histochemical Journal, 31, 1, 19-28, 1999.
(要約)
The granular convoluted tubule of the mouse submandibular gland contains a wide variety of biologically active proteins, including several kallikreins. The tubule is under multihormonal regulation, and is sexually dimorphic, being larger in males than in females. Correspondingly, levels of its various protein secretory products are more abundant in males than in females. However, isoelectric focussing studies show that the true tissue kallikrein, mK1, is more abundant in the female than in the male submandibular gland. In this study, an antiserum was prepared with restricted immunoreactivity for mouse mK1, and possibly other kallikrein family members of low abundance in the mouse submandibular gland, and used for the immunocytochemical staining of the granular convoluted tubule cells in the submandibular gland of adult male and female mice, by indirect enzyme-labeled and immunogold-labeled antibody methods for light and electron microscopy, respectively. The distribution of immunoreactive tubule cells showed an unusual sexual dimorphism. In males only a few scattered slender tubule cells were strongly stained, while the more typical large tubule cells were only occasionally weakly positive, and many of them were not stained. By contrast, in females slender tubule cells were not seen, and about two thirds of the more typical tubule cells showed moderate to strong immunostaining. Immunoelectron microscopy revealed that immunostaining was confined to the secretion granules in granular convoluted tubule cells in both sexes. The slender tubule cells of males had many strongly stained small apical secretion granules and occasional basal infoldings; in the weakly positive larger more typical tubule cells not all secretion granules were positive, and there was intergranular variation in the intensity of staining of positive granules. In females, although more tubule cells were stained, intergranular variations in staining intensity were also noted. In both sexes, many tubule cells did not contain any secretion granules that showed immunogold labeling for kallikreins. These findings establish that, in contrast to the situation for the majority of granular convoluted tubules proteins, mK1 and possibly other minor kallikrein family members are more abundant in the granular convoluted tubules of female mice, and that there is considerable variation in the content of these kallikreins not only between different tubule cells, but also in individual secretion granules in any given tubule cell in either sex.
(キーワード)
Animals / Antibodies / Antibody Specificity / Female / Immune Sera / Immunohistochemistry / Kallikreins / Male / Mice / Mice, Inbred ICR / Microscopy / Microscopy, Electron / Sex Characteristics / Sex Factors / Staining and Labeling / Submandibular Gland
Norihiko Maeda, Koichi Ogata, Setsuko Suemune, Yuji Yoshiko, Shumpei Niida, Mitsuteru Hosoi, Keita Hirano, Kazuo Hosoi and Katsunobu Miyata : Co-localization of estrogen and androgen receptors immunoreactivities in the trigeminal motor nucleus, Biomedical Research (India), 9, 3, 187-193, 1998.
49.
Kazuo Hosoi, Jun Tada, Keiko Tsumura, Norio Kanamori and Naoki Yamanaka : Expression of an alloenzyme of prorenin-converting enzyme in the submandibular gland of DBA/2N mice., The Journal of Biochemistry, 124, 2, 368-376, 1998.
Yamato Kikkawa, Naoki Yamanaka, Jun Tada, Norio Kanamori, Keiko Tsumura and Kazuo Hosoi : Prorenin processing and restricted endoproteolysis by mouse tissue kallikrein family ezymes (mK1, mK2, mK13, and mK22)., Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1382, 1, 55-64, 1998.
Kinji Kurihara, Shichiro Maruyama, Kazuo Hosoi, Seiichi Sato, Takao Ueha and Edward W Gresik : Regulation of Na+,K+-ATPase in submandibular glands of hypophysectomized male mice by steroid and thyroid hormones, The Journal of Histochemistry and Cytochemistry, 44, 7, 703-711, 1996.
(要約)
The effects of thyroid hormone, androgen, glucocorticoid, and mineralocorticoid on Na+,K+-ATPase activity and on levels of its alpha-subunit protein (alpha 1 isoform) in mouse submandibular gland (SMG) were studied by enzyme assay for ouabain-sensitive ATP hydrolysis, by quantitative densitometric scanning of Western blots, and by immunohistochemistry. To define the specific regulatory effects of various pituitary-dependent hormones on expression of Na+,K+-ATPase in the SMG, we treated hypophysectomized (hypox) male mice with triiodo-L-thyronine (T3), 5 alpha-dihydrotestosterone (DHT), dexamethasone (Dex), and aldosterone (Ald), injected singly or in combination. Na+,K+-ATPase was confined to the duct system of the SMG. In intact mice there was a gender difference in SMG Na+,K+-ATPase, with levels of the enzyme's activity and of its alpha 1-subunit being less in the glands of males. In males, hypophysectomy caused a rise in levels of Na+,K+-ATPase activity and in levels of the alpha 1-subunit protein of this enzyme, and in intensity of immunocytochemical staining for this subunit but there were no such changes in the SMG of hypox females. Changes caused by hormonal replacement to hypox males in Na+,K-ATPase activity, levels of its alpha 1-subunit, or the intensity of immunocytochemical staining for this subunit were complex. Ald had no effect. T3 or dexamethasone, given alone, induced Na+,K+-ATPase activity above control values (hypox males) and increased levels of its alpha 1-subunit protein and immunohistochemical staining for this subunit. By contrast, DHT did not cause a decline in any of these parameters. However, when treatment with T3 was combined with administration of Dex or DHT, enzymatic activity of Na+,K+-ATPase decreased but levels of the alpha 1-subunit protein and immunohistochemical staining for this subunit increased. Therefore, inductions of the alpha 1-subunit of this enzyme are not always correlated with increases in levels of activity of Na+,K+-ATPase, and we propose that both enzymatic and immunochemical analyses are essential for evaluation of hormonal regulation of Na+,K+-ATPase in salivary gland and in other tissues.
Yoshifumi Tajima, Kohtaro Kato, Shichiro Maruyama and Kazuo Hosoi : In vivo modulation of proliferating cell nuclear antigen in growth plate chondrocytes from normal, hypophysectomized, and growth hormone-treated hypophysectomized rats -A comparative immunohistochemical study with image analysis-, The Journal of Histochemistry and Cytochemistry, 44, 7, 713-720, 1996.
(要約)
Growth hormone (GH) regulates the proliferation and maturation of chondrocytes in the epiphyseal growth plate, in which a temporal dimension is superimposed on the septal organization of the tissue. In this study we investigated the in vivo effects of hypophysectomy (Hypox) and injection of GH into Hypox animals (Hypox + GH) on the proliferative activity of the growth plate chondrocytes. We assessed the immunohistochemical expression of proliferating cell nuclear antigen (PCNA) in paraffin-embedded tissues, using monoclonal antibody PC 10 against PCNA combined with immunogold-silver staining. We subjected the immunostained sections to computer-based image analysis by ACAS 570 interactive laser cytometry employing a conventional microscopic light source. Hypox was carried out on 20 rats at 8 weeks of age, half of which received a hypodermic injection of human GH at a dose of 1 IU/kg twice a day for 1 week after the operation. Another group of five rats of the same age were used as normal controls. In normal rats, a distinct PCNA immunoreaction was observed in the proliferative zone, whereas a remarkable diminution of the number of immunoreactive cells in this zone was apparent in Hypox animals. On the other hand, marked hyperplasia of PCNA-positive cells was seen in the proliferative zone of the Hypox + GH rat growth plate. The immunoreactive cells of this zone in Hypox + GH animals exhibited increased nuclear size and staining intensity of PCNA compared with those in normal and Hypox rats. These findings were further confirmed by computer-based image analysis of the specimens in terms of cell integrated value, area, perimeter, and shape factor. These different patterns of PCNA expression may imply that GH specifically promotes the proliferation phase of the chondrocytes in the proliferative zone. The data also suggest that GH influences not only cell replication activity but also cell kinetics of chondrocytes in the growth plate during their lifespan.
Kazuo Hosoi, Yoshimi Shioda, Takao Ueha, Toshiko Atsumi, Kenji Sugita and Kinji Kurihara : ATP- and EGF-stimulated phosphatidylinositol synthesis by two different pathways, phospholipase D and diacylglycerol kinase, in A-431 epidermoid carcinoma cells, Biochemistry and Cell Biology, 74, 2, 197-209, 1996.
54.
栗原 琴二, 丸山 七郎, 細井 和雄, Edward W Gresik, 上羽 隆夫 : 脳下垂体摘出マウス顎下腺のNa+, K+-ATPase活性およびその局在性に及ぼす甲状腺ホルモン並びにステロイドホルモンの影響, 日本唾液腺学会誌, 36, 41-43, 1995年.
Kenji Sugita, Kinji Kurihara, Kazuo Hosoi, Toshiko Atsumi, Tsutomu Takahashi, Minoru Kohno and Takao Ueha : Effects of pertussis toxin on signal transduction via P2-purinergic receptors in A-431 human epidermoidal carcinoma cells, Enzyme & Protein, 48, 4, 222-228, 1995.
(要約)
In A-431 cells, stimulation of P2-purinergic receptors with extracellular ATP caused production of inositol 1,4,5-trisphosphate (InsP3), followed by mobilization of Ca2+ from intracellular stores; Ca2+ influx from the extracellular fluid and breakdown of phosphatidylcholine (PtdCho) also accompanied this InsP3/Ca2+ signalling. When A-431 cells were pretreated with pertussis toxin (PTX), production of InsP3 and elevation of cytosolic Ca2+ were strongly inhibited. PTX also inhibited the Ca2+ influx, but the effect was much weaker than that for InsP3/Ca2+ elevation. No inhibitory effect was observed in ATP-stimulated PtdCho breakdown. These results suggest that there is a system(s) which mediates the functions of P2-purinergic receptors in addition to PTX-sensitive G-proteins.
Hiroshi Kamioka, Yoshiki Miki, Koji Sumitani, Kahori Tagami, Kunihiro Terai, Kazuo Hosoi and Terushige Kawata : Extracellular calcium causes the release of calcium from intracellular stores in chick osteocytes, Biochemical and Biophysical Research Communications, 212, 2, 692-696, 1995.
(要約)
We have recently demonstrated that a rise in the extracellular divalent cation concentration induces a rapid elevation of cytosolic calcium in chick osteocytes. Here, we demonstrate that cytosolic calcium elevation that occurs in osteocytes on exposure to elevated extracellular calcium is independent of membrane voltage and is insensitive to modulation by organic calcium channel modulators, namely, BAY K 8644, nicardipine, and nifedipine. However, the calcium elevation was sensitive to modulation by an intracellular calcium antagonist, TMB-8, suggesting that the cytosolic calcium elevation was due to mobilization of this cation from an intracellular store.
Jianxiang Qu, Kazuo Hosoi, Takahiro Shimojima, Tsuyoshi Oi and Katsumi Ikeda : Effects of FMLP and LPS on [Ca2+]i of peritoneal exudate polymorphonuclear leukocytes following onset of inflammation, Journal of Periodontal Research, 30, 3, 153-158, 1995.
(要約)
Because a general study of activated neutrophils may have relevance to periodontal diseases and accompanying inflammation, we studied a function of mouse polymorphonuclear leukocytes (PMNs) that exude into the peritoneal cavity in response to inflammation caused by i.p. injection of 2% casein. The effects of E. coli-lipopolysaccharide (E-LPS) and a chemotactic factor, N-formyl-N-methionyl-N-leucyl-L-phenylalanine (FMLP), on the level of intracellular calcium ([Ca2+]i) in these PMNs were examined. From analysis made with a laser cytometer (ACAS 570), the PMNs in exudates harvested 3-9 h after the onset of inflammation were shown to undergo [Ca2+]i elevation in response to 10(-6) M FMLP. The peak concentration of [Ca2+]i elicited by FMLP was highest in exudate cells 6 h after casein injection. In addition, about 65% of the PMNs in the 3-h exudate were FMLP sensitive displaying an elevated [Ca2+]i, whereas more than 85% of them in 6- and 9-h exudates became FMLP sensitive. Also, the maximum level of [Ca2+]i after FMLP stimulation was potentiated by pretreatment of the cells with E-LPS (0.2 microgram/ml). The present study suggests that PMNs induced by casein injection and appearing in mouse peritoneal exudate at different times possess significantly different ability to undergo [Ca2+]i elevation, and different susceptibility toward a chemotactic factor, FMLP.
Yoshifumi Tajima, Kohtaro Kato, Nobuo Utsumi and Kazuo Hosoi : Computer-aided image analysis applied to immunogold-silver staining: evaluation of proliferating cell nuclear antigen (PCNA)-reactive sites in paraffin sections, Histochemistry, 102, 3, 177-181, 1994.
(要約)
Feasibility of the combination of the immunogold-silver staining method (IGSS) and computer-aided image analysis was assessed for the detection of antigen in an immunostained, paraffin-embedded section. Using low-temperature IGSS, we stained a specimen of human oral squamous cell carcinoma with a monoclonal antibody, PC 10, against a proliferating cell nuclear antigen (PCNA/cyclin), and the section was analyzed by ACAS 570 interactive laser cytometry. The PCNA-positive cells, exhibiting a heteromorphic texture, were contrasted by the dark staining of their nuclei, but showed heterogeneity in staining intensity from cell to cell. Using a conventional microscope light source rather than a laser, and by employing the COMPLEMENT DATA program (which permits inversion of the data values) installed in the ACAS 570 software system, we were able to obtain a 'complemental image' which replicated the real immunohisto-morphology. Approximately 30-35 cells from three different areas in the same section were selected by DEFINE CELL and MARK AREA programs, and quantitative image analysis was performed in terms of cell integrated value, area, perimeter, and shape factor indicated in histogram form. The combined utilization of IGSS with computer-aided image analysis was demonstrated to offer a crucial advantage for the quantitative assessment of immunostained sections.
Toshiko Atsumi, Kazuo Hosoi and Takao Ueha : Involvement of high-affinity binding site for EGF receptor in formation of rounding in A-431 epidermoid carcinoma cells, Hormone and Metabolic Research, 26, 3, 141-144, 1994.
(要約)
The introduction of a bacterial aminoglycoside phosphotransferase gene (neo gene) into A-431 cells was found to result in disappearance of high-affinity binding sites of the epidermal growth factor receptor (EGFR), probably by affecting the phosphorylation level of the receptors. Using A-431 cells and their neo gene-transfectants, we studied the relation between "rounding" and the high-affinity sites for EGF; and we also examined the role of protein kinase C (PKC) and A (PKA) in the EGF-induced cell rounding. Pretreatment of A-431 and their transfectant cells with 12-O-tetradecanoylphorbol 13-acetate (TPA; 100 ng/ml), an activator of PKC, for 30 min inhibited both the EGF-induced cell rounding and expression of high-affinity binding sites for EGF. However, both of these responses were recovered when cells were pretreated with TPA for 20 h, which treatment is known to result in depletion of PKC by a process called "down regulation". A similar recovery was also observed when cells were pretreated with forskolin (100 microM), an activator of PKA, for 30 min. Both cell rounding and EGFR high-affinity binding sites disappeared by activation of PKC, and reappeared by activation of PKA. These results suggest that the rounding of A-431 cells by EGF was induced via the high-affinity binding sites of EGFR.
Kazuo Hosoi, Susumu Tsunasawa, Kinji Kurihara, Hideyuki Aoyama, Takao Ueha, Toyoaki Murai and Fumio Sakiyama : Identification of mK1, a true tissue (glandular) kallikrein of mouse submandibular gland: Tissue distribution and a comparison of kinin-releasing activity with other submandibular kallikreins, The Journal of Biochemistry, 115, 1, 137-143, 1994.
(要約)
The protein structure, kinin-releasing activity, and tissue distribution of four major proteinases of mouse submandibular gland (mK22, mK9, proteinase F, proteinase P) were studied. When compared with the deduced amino acid sequence of each member of the tissue (glandular) kallikrein gene family, the amino acid sequence of proteinase F determined (approximately 40% of the total) was found to agree completely with the deduced amino acid sequence of mKlk-1. The proteinase P sequence, on the other hand, agreed with that of the product of mKlk-13, mK13 (prorenin-converting enzyme). Proteinase F had the strongest kininogenase activity for both low-molecular-weight and high-molecular-weight kininogen, while mK22 had 1/6 and 1/50 the activity of proteinase F for the respective kininogen substrate. Kininogenase activities of mK9 and proteinase P were less than 1/100 of the activity of proteinase F for both substrates. Acting on the two kininogen substrates, kallikreins mK22, mK9, and proteinase F, but not proteinase P, specifically released bradykinin, suggesting that the former three kallikreins strictly recognized peptide sequences around bradykinin in these substrate molecules but proteinase P recognized several sites in these molecules. Significant amounts of proteinase F, but not mK22 and others, were present in the urine, pancreas and digestive organs, as well as in the salivary glands. The present results revealed that the former proteinase F is identical to mK1, tissue/renal kallikrein, and confirmed its characteristics as a true kallikrein on the basis of its kinin-releasing activity and tissue distribution.
Kinji Kurihara, Kazuo Hosoi, Takao Ueha, Nobuo Nakanishi and Shozo Yamada : Effects of nerve growth factor and dexamethasone on Na+,K+-ATPase of cultured PC12 cells, Hormone and Metabolic Research, 26, 1, 14-18, 1994.
(要約)
When PC12h cells were cultured for 4 days in the presence of 50 ng/ml of nerve growth factor (NGF), they showed elongated dendrites and specifically increased Na+,K(+)-ATPase activity. Either singly or in combination with NGF, dexamethasone also increased the specific activity of this enzyme. Western blot analysis using anti-alpha 1 and anti-alpha 2 antisera showed that PC12h cells, either before or after hormone treatment, contained the alpha 1 isoform but not the alpha 2 one. We conclude, therefore, that NGF induces Na+,K(+)-ATPase concomitantly with neuronal differentiation in PC12h cells but that the growth factor does not induce formation of the myelin sheath, which normally expresses the alpha 2 isoform of Na+,K(+)-ATPase.
Kazuo Hosoi, Kinji Kurihara and Takao Ueha : Bradykinin-stimulated transient modulation of epidermal growth factor receptors in A-431 human epidermoid carcinoma cells, Journal of Cellular Physiology, 157, 1, 1-12, 1993.
(要約)
Of nine biological factors (ATP, bradykinin, vasopressin, substance P, angiotensin II, norepinephrine, epinephrine, 12-tetradecanoylphorbol 13-acetate (TPA), and A23187 calcium ionophore) examined, bradykinin, as well as ATP, TPA, and A23187, significantly increased the phosphorylation of epidermal growth factor (EGF) receptors and reduced the binding of EGF to their high-affinity site. The reduction in EGF binding by bradykinin, ATP, and TPA was similarly reversed by concomitant incubation with staurosporine, a protein kinase C inhibitor, implying that the phosphorylation of EGF receptors was catalyzed probably by a protein kinase C of the same or similar type in each case. This possibility was confirmed by the fact that the major phosphorylation site of EGF receptors by the stimulation with either bradykinin, ATP, or TPA was the same (Thr-654). Different from the stimulations with ATP and TPA, the effect of bradykinin of decreasing the high-affinity EGF binding was transient (a minimum binding at 2.5 min); the reduced EGF binding was, however, sustained for up to 30 min in the presence of calyculin A, a phosphoprotein phosphatase inhibitor. Moreover, the homogenate prepared from bradykinin-stimulated A-431 cells had stronger dephosphorylation activity for phosphorylated EGF receptors than that from control cells. These results suggest that bradykinin stimulates both the protein kinase C system and a phosphoprotein phosphatase(s) activity in A-431 cells. Such biphasic effects of bradykinin to phosphorylate and dephosphorylate EGF receptors via protein kinase C and a phosphoprotein phosphatase, respectively, imply a homeostatic control of receptor function in regulating phosphorylation level by the same bioactive factor.
Shichiro Maruyama, Kazuo Hosoi, Takao Ueha, Masamichi Tajima, Seiichi Sato and Edward W Gresik : Effects of female hormones and 3,5,3'-triiodothyronine or dexamethasone on induction of epidermal growth factor and proteinases F, D, A, and P in the submandibular glands of hypophysectomized male mice, Endocrinology, 133, 3, 1051-1060, 1993.
(要約)
The effects of progesterone (Pro), 5 alpha-dihydrotestosterone (DHT), 17 beta-estradiol (E2), T3, and dexamethasone (Dex), given alone or in combination, on induction of epidermal growth factor (EGF) and proteinase isozymes in the submandibular glands of hypophysectomized mice were examined. Each hormone, except E2, acting alone had essentially comparable inductive effects on EGF concentrations and on total proteinase activity. E2 alone had no inductive effect at all. Pro acted synergistically with T3, but its inductive effect was diminished when given with Dex. E2 was not synergistic with T2, and it inhibited the effect of Dex; it also partially blocked the action of DHT on induction of proteinase activity but not of EGF. Simultaneous administration of all five hormones restored total proteinase activity completely but EGF levels to only 50% of values for intact males, respectively. Submandibular proteinases were resolved by isoelectric focusing into four isozymes: proteinase F (pI 4.8), proteinase D (pI 5.8), proteinase A (pI 6.2), and proteinase P (pI 10.0). Pro alone slightly increased levels of proteinase F but greatly raised levels of the other three isozymes. This inductive action was augmented when Pro was given with T3 but blocked when it was given with Dex. E2 alone not only failed to induce any of the isozymes, but even further reduced levels of proteinase F. It also decreased the inductive effects of T3 on these isozymes, and with Dex completely blocked induction of proteinases D, A, and P. E2 plus DHT suppressed proteinase F levels and only induced proteinase A. All five hormones together reestablished the isozyme profile seen in intact males. These results show that Pro by itself is as capable as androgens, thyroid hormone, or glucocorticoid in regulating expression of these submandibular polypeptides, and that its action can be modulated by other pituitary-dependent hormones. In addition, they demonstrate that E2 does not regulate their expression, and that it has an inhibitory effect on the inductive action of other hormones. Last, they indicate that these various hormones may regulate expression of EGF and of each of the individual proteinase isozymes differently.
丸山 七朗, 細井 和雄, 上羽 隆夫, Edward W Gresik, 田島 雅道, 佐藤 精一 : 脳下垂体除去マウス顎下腺の各種proteinase (F, D, A, P)及びEGFに対する女性ホルモンと副腎皮質及び甲状腺ホルモンの併用による動態, 日本唾液腺学会誌, 33, 35-39, 1992年.
67.
Kinji Kurihara, Kazuo Hosoi and Takao Ueha : Characterization of ecto-nucleoside triphosphatase on A-431 human epidermoidal carcinoma cells, Enzyme, 46, 4,5, 213-220, 1992.
68.
Kazuo Hosoi, Mariko Fujishita, Kenji Sugita, Kinji Kurihara and Takao Ueha : P2-Purinergic receptors and cellular calcium metabolism in A 431 human epidermoid carcinoma cells, American Journal of Physiology, Cell Physiology, 262, 31, 635-643, 1992.
69.
Kazuo Hosoi, Shichiro Maruyama, Takao Ueha, Seiichi Sato and Edward W Gresik : Additive and/or synergistic effects of 5a-dihydrotestosterone, dexamethasone, and triiodo-L-thyronine on induction of proteinases and epidermal growth factor in the submandibular gland of hypophysectomized mice, Endocrinology, 130, 2, 1044-1055, 1992.
Kazuo Hosoi, Kenji Sugita, Yoshimi Shioda, Akiko Kodama and Takao Ueha : Involvement of cellular calcium incorporated from the medium in production of inositol trisphosphate in A431 cells, Cellular Signalling, 3, 1, 79-84, 1991.
72.
Kenji Sugita, Kazuo Hosoi, Yoshimi Shioda and Takao Ueha : Effects of Ca2+ and Mg2+ on ATP-dependent 45Ca2+ influx in A-431 human epidermoidal carcinoma cells, Biochemistry and Cell Biology, 69, 1, 29-35, 1991.
73.
Kazuo Hosoi, Takao Ueha, Hidekazu Fukuuchi, Minoru Kohno and Tsutomu Takahashi : Reversal of relative proteinase F activity and onset of androgen-dependent proteinases in the submandibular gland of postnatal mice, Biochemistry International, 22, 1, 179-187, 1990.
74.
Kinji Kurihara, Kazuo Hosoi, Akiko Kodama and Takao Ueha : A new electrophoretic variant of a subunit of Na+,K+-ATPase from the submandibular gland of rats, Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1039, 2, 234-240, 1990.
(要約)
The alpha catalytic subunits of Na+/K(+)-ATPase were isolated from the kidney and brain of rats (alpha 1 and alpha 2, respectively). The antisera raised against these subunits were used as probes to analyze the isoform of catalytic subunits of Na+/K(+)-ATPase in various tissues of rats. Of 27 rat tissues examined, most had a catalytic subunit identical to alpha 1 but some, such as the nervous and muscle tissues, had both alpha 1 and alpha 2 isoforms as judged by their reactivities to antisera and their electrophoretic mobility. We found that the submandibular gland contained a new electrophoretic variant of immunoreactive alpha subunit (designated alpha(S) in this report) in addition to alpha 1 identical to those found in kidney and brain. The new variant, alpha(S), strongly cross-reacted with anti-alpha 1 antiserum, but to a lesser extent with anti-alpha 2 antiserum. The alpha(S) had a molecular mass which was found to be slightly less (approx. 90 kDa) than brain and kidney alpha 1. We examined whether or not the alpha(S) is formed by proteolytic cleavage of alpha subunits during preparation and concluded that this is not the case. The alpha(S) reacted with [gamma-32P]ATP, resulting in the formation of radioactive alpha subunit which was stabilized by 2 mM ouabain but which was labile in the presence of 70 mM potassium chloride. Since N-terminal amino acid sequence of alpha(S) protein [G()DKY()PAAVS] corresponds exactly and uniquely with the sequence of the alpha 1 chain between residues 1 and 11, it is very probable that alpha(S) protein originated from alpha 1 protein following the post-translational processing.
Kazuo Hosoi, Toshiko Atsumi and Takao Ueha : A human-mouse hybrid cell line expressing both HLA and H-2 antigens, The Japanese Journal of Physiology, 40, 2, 297-304, 1990.
Kazuo Hosoi and Michael Edidin : Exogenous ATP and other nucleoside phosphates modulate epidermal growth factor receptors of A-431 epidermoid carcinoma cells, The Proceedings of The National Academy of Sciences USA, 86, 12, 4510-4514, 1989.
78.
Kazuo Hosoi, Kinji Kurihara, Akiko Kodama, Yoshimi Shioda, Kenji Sugita and Takao Ueha : Postnatal changes in an a-subunit isoform, a(S), of Na+,K+-ATPase in the submandibular gland of rats, Enzyme, 42, 3, 152-159, 1989.
79.
Kazuo Hosoi, Yoshimi Shioda, Akiko Kodama, Kenji Sugita, Kinji Kurihara, Toyoaki Murai, Akira Nemoto, Toshiko Atsumi and Takao Ueha : A simple and rapid purification procedure for the tetramethylrhodamine-labeled antibodies, The Japanese Journal of Physiology, 39, 2, 317-324, 1989.
(要約)
A conjugate preparation of antibody (Fab) and tetramethylrhodamine (TMR) generally adsorbs free TMR which is very difficult to remove because of its strong hydrophobic binding. On the basis of criteria such as sodium dodecyl sulfate (SDS) gel electrophoresis and staining of the plasma membrane of live cells, we found that simple extraction with n-butyl alcohol or iso-amyl alcohol could remove the contaminating free dye. The procedure is especially useful when one needs to prepare conjugates with low nonspecific binding for the study of lateral diffusion of cell membrane-associated antigens.
(キーワード)
1-Butanol / Butanols / Carcinoma, Squamous Cell / Cell Line / Electrophoresis, Polyacrylamide Gel / Immunoglobulin Fab Fragments / Pentanols / Rhodamines / Xanthenes
(文献検索サイトへのリンク)
● PubMed @ National Institutes of Health, US National Library of Medicine (PMID): 2761125
Kazuo Hosoi, Dilip S. Kittur and Michael Edidin : Modulation of HLA antigens in response to the binding of epidermal growth factor by A431 cells, FEBS Letters, 231, 2, 371-377, 1988.
81.
Toyoaki Murai, Kinji Kurihara, Kenji Sugita, Kazuo Hosoi and Takao Ueha : Androgen responsiveness of proteinase F in the submandibular salivary glands of inbred mice, The Japanese Journal of Physiology, 37, 5, 871-879, 1987.
(要約)
Using the radioimmunoassay technique, we studied androgenic regulations of proteinase F in the submandibular salivary glands (SMG) of various mouse strains (ICR, BALB/CA, C3H/HeN, C57BL/10N, and DBA/2N). The proteinase contents in BALB/CA and DBA/2N mice were almost the same between males and females. However, prominent sex differences in enzyme content were seen in all other strains; i.e., male glands of C3H/HeN and C57BL/10N contained more proteinase F than female glands did. On the contrary, the enzyme level in the ICR was exceedingly higher in females than in males. The enzyme contents in ICR SMG increased in males following castration and decreased after androgen injection in both castrated males and normal females. On the contrary, the enzyme level in C3H/HeN was decreased by castration and increased by androgen injection. Therefore, these sex differences in ICR and C3H/HeN mice depend on androgen, though the direction of the response is opposite.
Shigeyasu Tanaka, Kazuo Hosoi, Izumi Tanaka, Masayoshi Kumegawa and Takao Ueha : Immunocytochemical study of proteinase F in the mouse submandibular gland, The Journal of Histochemistry and Cytochemistry, 32, 6, 585-592, 1984.
83.
Kazuo Hosoi, Izumi Tanaka, Toyoaki Murai and Takao Ueha : Inhibitory effect of androgen on the synthesis of proteinase F in the male mouse submandibular gland, The Journal of Endocrinology, 100, 2, 253-262, 1984.
84.
Kazuo Hosoi, Izumi Tanaka, Yukihito Ishii and Takao Ueha : A new esteroproteinase (proteinase F) from the submandibular gland of female mice, Biochimica et Biophysica Acta (BBA) - General Subjects, 756, 2, 163-170, 1983.
(要約)
One of the esteroproteinases present in the submandibular glands of female mice was purified and characterized. The enzyme, designated proteinase F in this report, had a pI value of 4.6 and a molecular weight of 27600, being comprised of two subunits of 10000 and 18000 daltons. The amino acid composition of proteinase F resembled that of the epidermal growth factor-binding protein, but antiserum against proteinase F only reacted weakly against the binding protein. Proteinase F had an optimum pH at around 9.0 and was strongly inhibited by Cu2+ and Hg2+ (42 and 76% inhibition, respectively, at a concentration of 4 x 10(-6) M). It was also inhibited by aprotinin, phenylmethylsulfonylfluoride, iodoacetamide, leupeptin, antipain, and benzamidine but neither by trypsin inhibitors from pancrease, soybean, or ovomucoid, nor by TLCK, TPCK, and epsilon-amino-n-caproic acid. Although its actual physiological function has yet to be determined, these properties indicate that proteinase F is a new enzyme, being distinguished from known proteinases, kallikrein, plasmin, trypsin, chymotrypsin, tonin, angiotensin-converting enzyme, proteinase A (beta-nerve growth factor endopeptidase), proteinase D (epidermal growth factor-binding protein), P-esterase, renin A, and renin C. Proteinase F was present in the submandibular glands of female mice more abundantly than in those of males, but it increased in males following castration. Thus, proteinase F appears to be affected by male hormones in vivo.
Kazuo Hosoi, Shizuko Kamiyama, Toshiko Atsumi, Akira Nemoto, Izumi Tanaka and Takao Ueha : Characterization of two esteroproteases from the male mouse submandibular gland, Archives of Oral Biology, 28, 1, 5-11, 1983.
86.
Kazuo Hosoi, Izumi Tanaka and Takao Ueha : Induction of epidermal growth factor by tri-iodo-L-thyronine in the submandibular glands of mice with testicular feminization, The Journal of Biochemistry, 90, 1, 267-270, 1981.
87.
Kazuo Hosoi, Tadayoshi Kida and Takao Ueha : Induction of various androgen-dependent esteroproteases (trypsin-like and chymotrypsin-like enzymes) by tri-iodo-L-thyronine in the submandibular glands of female mice and mice with testicular feminization, The Journal of Biochemistry, 89, 6, 1793-1798, 1981.
88.
Shizuko Kobayashi, Kazuo Hosoi and Takao Ueha : Purification and properties of b-N-acetylhexosaminidase from mouse submandibular gland, Archives of Oral Biology, 25, 11,12, 753-758, 1980.
89.
Masahiko Hiramatsu, Keiko Hatakeyama, Kazuo Hosoi and Naomi Minami : Effect of autonomic agents on the secretion of N-acetyl b-glucuronidase of mouse submaxillary gland, Journal of Dental Research, 59, 8, 1439-1441, 1980.
90.
Kazuo Hosoi, Shizuko Kobayashi, Takao Ueha, Shichiro Maruyama, Seiichi Sato, Taishin Takuma and Masayoshi Kumegawa : Induction of androgen-dependent protease and serous-like granules by tri-iodothyronine in the submandibular gland of mice with testicular feminization, The Journal of Endocrinology, 83, 3, 429-434, 1979.
91.
Masahiko Hiramatsu, Keiko Hatakeyama, Kazuo Hosoi and Naomi Minami : Regulation of delayed-hypersensitivity response by the submandibular gland of male mice, Immunology, 37, 4, 869-872, 1979.
92.
Kazuo Hosoi, Shizuko Kobayashi, Masahiko Hiramatsu, Naomi Minami and Takao Ueha : Androgenic regulation of N-acetyl b-glucuronidase activity in the submandibular glands of mice, The Journal of Biochemistry, 85, 6, 1483-1488, 1979.
93.
Kazuo Hosoi, Shizuko Kobayashi and Takao Ueha : Affinity adsorption of L-glutamine D-fructose-6-phosphate aminotransferase to Sepharose coupled with p-chloromercuribenzoate, Biochemical and Biophysical Research Communications, 85, 2, 558-563, 1978.
94.
Kazuo Hosoi, Shizuko Kobayashi and Takao Ueha : Sex difference in L-glutamine D-fructose-6-phosphate aminotransferase activity of mouse submandibular gland, Biochimica et Biophysica Acta (BBA) - General Subjects, 543, 3, 283-292, 1978.
(要約)
L-Glutamine D-fructose-6-phosphate aminotransferase (2-amino-2-deoxy-D-glucose-6-phosphate ketol-isomerase (amino-transferring), EC 5.3.1.19) activities in the three main salivary glands of male and female mice were measured. It was found that the activity in the submandibular gland was about 10 times more in females than in males, whereas the activities in the sublingual and parotid glands of males and females were similar. The activity in the submandibular gland of female mice was not affected appreciably by ovariectomy but it decreased to the level in males on injection of testosterone. The activity in males was not affected appreciably by injection of progesterone or 17beta-estradiol, but it increased to the level in females after castration. The increased activity in castrated male mice was decreased again to the normal level by testosterone injection. Thus, this sex difference is caused by androgen, not by female hormones. On the basis of in vivo experiments using actinomycin D, it was suggested that testosterone produced an "enzyme inhibitor", which suppressed the enzyme activity in the submandibular glands of androgen-rich animals.
Kazuo Hosoi, Shizuko Kobayashi and Takao Ueha : Influence of androgen on b-glucuronidase activity in liver, kidney and submandibular salivary gland of the mouse, Archives of Oral Biology, 23, 10, 905-909, 1978.
96.
Kazuo Hosoi, Kazushi Aoyama and Takao Ueha : Effect of autonomic agents on the amount of androgen-dependent granules in convoluted tubular cells of the mouse submandibular gland, Canadian Journal of Physiology and Pharmacology, 56, 4, 634-641, 1978.
97.
Kazuo Hosoi, Kazushi Aoyama and Takao Ueha : Regulation of the secretory process of granular components from the convoluted tubular cells of the mouse submandibular gland, Journal of Dental Research, 57, 1, 87-90, 1978.
Kazuo Hosoi and Takao Ueha : Effects of sex hormones on synthesis of proteins contained in granules present in convoluted tubular cells of mouse submandibular glands, The Journal of Biochemistry, 82, 2, 351-358, 1977.
100.
Kazuo Hosoi and Toshikazu Nakamura : Effect of testosterone on the amount of serous-like granules in convoluted tubular cells of mouse submandibular glands, The Journal of Biochemistry, 81, 3, 739-748, 1977.
101.
Taishin Takuma, Toshikazu Nakamura, Kazuo Hosoi and Masayoshi Kumegawa : Binding protein for 5α-dihydrotestosterone in mouse submandibular gland, Biochimica et Biophysica Acta (BBA) - General Subjects, 496, 1, 175-181, 1977.
(要約)
The changes in the levels of the binding protein for 5 alpha-dihydrotestosterone in cytoplasmic extract of the submandibular glands during development were compared in male and female mice using a DEAE-cellulose filter assay. The binding protein was first detectable 5 days after birth in both sexes, at a time coincident with androgen-independent cytodifferentiation of the convoluted tubular cells in the submadibular gland. The level of the binding protein in female mice was maintained at 5 pmol/mg protein after birth, whereas in males it began to decrease from 3 weeks after birth with inccrease in serum testosterone, becoming much less than a quarter of the level in females or immature mice by 4 weeks after birth. However, after castration, the level of detectable binding protein in mature male mice increased within 7 days to the same level as that in females or immature mice. This suggests that the low binding capacity for exogenous hormone in mature male mice is due to occupancy of the binding sites by endogenous hormone.
Kazuo Hosoi, Gilbu Soe, Tomisaburo Kakuno and Takekazu Horio : Effects of pH indicators on various activities by chromatophores of Rhodospirillum rubrum, The Journal of Biochemistry, 78, 6, 1331-1346, 1975.
105.
Tatsuo Oku, Kazuo Hosoi, Gilbu Soe, Tomisaburo Kakuno and Takekazu Horio : Energization of NTP-NDP kinase by acid base transition leading to ATP formation, The Journal of Biochemistry, 76, 1, 233-235, 1974.
106.
Tomisaburo Kakuno, Kazuo Hosoi, Tomihiko Higuti and Takekazu Horio : Electron and proton transports in Rhodospirillum rubrum chromatophores, The Journal of Biochemistry, 74, 6, 1193-1203, 1973.
107.
Kazuo Hosoi, Setsuko Yoshimura, Gilbu Soe, Tomisaburo Kakuno and Takekazu Horio : Competition between Pi and pH indicators in photosynthetic ATP formation with chromatophores of Rhodospirillum rubrum, The Journal of Biochemistry, 74, 6, 1275-1278, 1973.
108.
Katsuzo Nishikawa, Kazuo Hosoi, Junnosuke Suzuki, Setsuko Yoshimura and Takekazu Horio : Formation and decomposition of pyrophosphate related to bacterial photophosphorylation, The Journal of Biochemistry, 73, 3, 537-553, 1973.
109.
Toshikazu Nakamura, Kazuo Hosoi, Katsuzo Nishikawa and Takekazu Horio : Effects of injection of chromatins from Rhodamine sarcoma into rats on pI-isozymes of liver pyruvate kinase, Gann : Japanese Journal of Cancer Research, 63, 2, 239-250, 1972.
Toshiko Atsumi, Kazuo Hosoi and Takao Ueha : Stability of HLA and H-2 expression on the surface of human-mouse hybrid cells prepared by cell hybridization technique, Journal of Meikai University, School of Dentistry, 20, 2, 158-169, 1991.
Kazuo Hosoi, Chenjuan Yao, Takahiro Hasegawa, Hiroshi Yoshimura and Tetsuya Akamatsu : Dynamics of Salivary Gland AQP5 under Normal and Pathologic Conditions, International Journal of Molecular Sciences, 21, 4, 1182, Feb. 2020.
Shingo Kurabuchi, Chenjuan Yao, Gang Chen and Kazuo Hosoi : Reversible Conversion among Subtypes of Salivary Gland Duct Cells as Identified by Production of a Variety of Bioactive Polypeptides, Acta Histochemica et Cytochemica, 52, 4, 59-65, Aug. 2019.
(要約)
Four major kallikreins (mK1, mK22, mK9, and mK13) were identified in the mouse submandibular gland (SMG). mK1, a true tissue kallikrein, was used as a protein marker to identify different types of SMG granular convoluted tubule (GCT) cells along with epidermal growth factor (EGF), nerve growth factor (NGF), and renin. Kallikrein mK1 was localized in a very small number (~5%) of GCT cells, which were scattered throughout the GCT, indicating that the majority of GCT cells are mK1-negative. Among mK1-positive cells, particularly strong signals were observed in a small number of narrow cells, recognized as slender granular cells (SG cells, Type IV), in the GCT. After postnatal development of the SMG, GCT cells are no longer uniform based on the bioactive substances (mK1, EGF, NGF, and renin) that they produce and secrete. GCT cells were classified into four subtypes, Types I-IV, and it became clear that these subtypes are complicatedly and reversibly converted by the endocrine hormones 5α-dihydrotestosterone (DHT) and triiodothyronine (T). Duct segments with similar morphology or hormone dependency were recognized in the sublingual and parotid glands. The presence of duct cells with such characteristics is therefore a common feature of the three major salivary glands of rodents.
Kazuo Hosoi, Javkhlan Purevjav and Chenjuan Yao : Possible Existence of a Salivary Gland-Oral Mucosa/Gingiva Axis Under Challenges by Endotoxins, Journal of Molecular and Cellular Biology Forecast, 1, 1, 1002, Jan. 2018.
5.
Kazuo Hosoi : Physiological role of aquaporin 5 in salivary glands, Pflügers Archiv : European Journal of Physiology, 468, 4, 519-539, Nov. 2015.
(要約)
Regarding the 13 known mammalian aquaporins (AQPs), their functions in their expressing tissues, effects of their mutation/polymorphisms in humans, and effects of knockout of their genes are summarized in this review article. The roles of AQP5, an exocrine gland-type water channel, in the salivary gland under normal and pathophysiological conditions are reviewed in detail. First, the involvement of AQP5 in water secretion from acinar cells was demonstrated by measuring volume changes of acini/acinar cells, as well as activation energy (E a) in transepithelial water movement by NMR spectrometry, and a functional linkage between AQP5 and TRPV4 was suggested. Next, involvement of the parasympathetic nervous system on the AQP5 levels in the acinar cells of the submandibular and that of a β-adrenergic agonist on those in the parotid gland are described. That is, chorda tympani denervation induces autophagy of the submandibular gland, causing AQP5 degradation/metabolism, whereas isoproterenol, a β-adrenergic agonist, causes first an increase then decrease in AQP5 levels in the parotid gland, which action is coupled with the secretory-restoration cycle of amylase-containing secretory granules. The PG also responded to endotoxin, a lipopolysaccharide that activates NF-κB and MAPK pathways. Elevated NF-κB and AP-1 (c-Fos/c-Jun) form a complex that can bind to the NF-κB-responsive element on the AQP5 promoter and thus potentially downregulate AQP5 transcription. Salivary gland pathologies and conditions involving AQP5 and possible treatments are described as well.
Most Parvin Nahid, Jun Tada, Tetsuya Akamatsu and Kazuo Hosoi : Structure, function, and transcriptional regulation of aquaporins, proteins of the water channel family, Dentistry in Japan, 37, 26-31, Mar. 2001.
12.
Edward W Gresik, Kazuo Hosoi, Kinji Kurihara, Shichiro Maruyama and Takao Ueha : The rodent granular convoluted tubule cell - An update, European Journal of Morphology, 34, 3, 221-224, Aug. 1996.
(要約)
The cells of the granular convoluted tubule (GCT) of the rodent submandibular gland (SMG) are under complex developmental and multihormonal regulation. Recent findings indicate that GCT cells also synthesize transforming growth factor alpha (TGF-alpha), hepatocyte growth factor, erythroid differentiation factor, endothelin, and insulin-like growth factor, as well as several novel androgen-dependent proteins of unknown function. The GCTs of hypophysectomized mice provide a convenient model to study multihormonal regulation of gene expression. The GCT system of the rodent SMG also is a fruitful model for study of hormone receptors.
Mileva Ratko Karabasil, Kwartarini Murdiastuti, Nunuk Purwanti, Azlina Ahmad, Javkhlan Purevjav, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : Effects of natural point mutation of rat aquaporin 5 expressed in vitro on its capacity of water permeability and membrane trafficking, The Journal of Medical Investigation : JMI, 56, Suppl, 398-400, Tokushima, Dec. 2009.
(要約)
In the colony of Sprague-Dawley (SD) strain, we found that there were rats expressing a mutant AQP5, which has a point mutation at nt 308 (G308A), leading to a replacement of (103)Gly with (103)Asp in the 3rd transmembrane domain. The mutant molecule scarcely expressed in the acinar cells, probably because of ineffective trafficking. The mutant molecule, however, showed normal water permeability when assessed by the oocyte system.
Purevjav Javkhlan, Yuka Hiroshima, Azlina Ahmad, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu, Jun-ichi Kido, Toshihiko Nagata and Kazuo Hosoi : Induction of calprotectin mRNAs by lipopolysaccharide in the salivary gland of mice, The Journal of Medical Investigation : JMI, 56, Suppl, 287-289, Tokushima, Dec. 2009.
(要約)
Calprotectin is a major cytosolic calcium-binding protein of leukocytes which belongs to the S100 protein family. S100A8 and S100A9, major types of calprotectin are heterodimeric complexes being composed of light- and heavy-chain subunits. The calprotectin levels in the plasma, feces, synovial fluid, gingival crevicular fluid, dental calculus and saliva change when the host animal suffers from several inflammatory diseases. Members of Toll-like receptor (TLR) family are pattern-recognition receptors for lipopolysaccharide (LPS) and other pathogens. Here we examined if the biological role of TLR receptor is reflected to the calprotectin expression in the salivary gland. Time course study by using real-time RT-PCR detected higher levels of S100A8 and S100A9 mRNA at 1.5-3 h after injection of LPS in both the submandibular gland (SMG) and parotid gland (PG) of C3H/HeN mice but not in the same tissues of C3H/HeJ, a TLR-4 mutant strain, indicating that this induction is mediated via the TLR-4. These results indicate that, an inflammatory marker, calprotectin, is expressed in the mouse salivary gland and that LPS stimulated its synthesis. Calprotectin (S100A8/A9) showed minimum expression in all cellular segments in the SMG except excretory duct cells, which showed strong signal at the cytoplasm. LPS induced their expressions in the granular convoluted tubular cells and striated duct cells. In the PG, these proteins were expressed very weakly in both duct and acinar cells with a little stronger staining for the former cells. LPS injection induced calprotectin (S100A8/A9) in both duct and acinar cells especially in the former cells.
Azlina Ahmad, Xuefei Li, Purevjav Javkhlan, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : Down-regulation of submandibular gland AQP5 following parasympathetic denervation in rats, The Journal of Medical Investigation : JMI, 56, Suppl, 273-276, Tokushima, Dec. 2009.
(要約)
Following chorda tympani denervation (CTD, parasympathetomy), the protein levels of aquaporin5 (AQP5) as well as AQP1 and Na(+)K(+)ATPase alpha-subunit in the rat submandibular gland (SMG) were found to be decreased significantly. However, the level of another membrane protein, dipeptidyl peptidase IV was not affected by CTD, suggesting a selective reduction of AQP5, AQP1, and Na(+)K(+)ATPase alpha -subunit proteins by CTD. However, the AQP5 mRNA level was scarcely affected by CTD, which suggested that transcription process of AQP5 was unaffected by this operation. AQP5 protein was shown to be degraded in vitro by the extract of the SMG obtained from normal rat; inhibitor experiments in vitro suggested cathepsin B was a responsible enzyme. Co-localization of AQP5 and LAMP-2, a lysosomal marker, implicated AQP5 is degraded in lysosomes. A significant increase in the protein levels of LC3-II, an autophagy marker, at day 1 after CTD, and co-localization of the LC3 protein and AQP5, suggested that CTD activated autophagy of SMG, leading to AQP5 degradation.
Nunuk Purwanti, Azlina Ahmad, Mileva Ratko Karabasil, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : Involvement of the IL-6/STAT3/Sca-1 system in proliferation of duct cells following duct ligation in the submandibular gland of mice, The Journal of Medical Investigation : JMI, 56, Suppl, 253-254, Tokushima, Dec. 2009.
(要約)
Ligation of the main excretory duct (MED) of the mouse submandibular gland (SMG) induced the expression of Sca-1, a stem cell marker. Sca-1 expression increased prominently in almost all of cells in the duct system, except the acinar cells. Sca-1 induction was accompanied with phosphorylated-STAT3 (Y705) elevation, which was localized in the nuclei of all duct cells. Electrophoretic mobility shift assay (EMSA) confirmed the specific binding of STAT3 to the GAS sequence, a biding site of gamma interferon activating site. Present study suggested one of the initial steps of the tissue regeneration after injury includes STAT3 pathway.
Tetsuya Akamatsu, Azlina Ahmad, Purevjav Javkhlan, Takahiro Hasegawa, Chenjuan Yao and Kazuo Hosoi : Salivary Gland Development: Its mediation by a subtilisin-like proprotein convertase, PACE4, The Journal of Medical Investigation : JMI, 56, Suppl, 241-246, Tokushima, Dec. 2009.
(要約)
The submandibular gland (SMG) develops under the epithelial-mesenchymal interaction. Its process is regulated by various growth/differentiation factors, which are synthesized as inactive precursors and activated via the limited proteolysis at their multi basic amino acid site(s) such as Arg-X-Lys/Arg-Arg. Although many of these processing steps are elucidated to be catalyzed by subtilisin-like proprotein convertases (SPCs), little is known about the role of SPCs in the SMG development. Here, we focused upon the physiological role of PACE4 (SPC4), a member of SPC family, in the SMG development. In the organ culture system of rat embryonic SMG (E15), Dec-RVKR-CMK, a potent inhibitor for SPCs, inhibited the salivary branching and the expression of an exocrine gland type water channel, AQP5. However, other peptidyl-CMKs and inhibitors for trypsin-like serine proteases including leupeptin did not affect the salivary branching and AQP5 expression. Dec-RVKR-CMK also suppressed the expression of PACE4, but not furin, another member of the family. The specific antibody for the catalytic domain of PACE4 suppressed the salivary branching and AQP5 expression similarly. These inhibitory effects of Dec-RVKR-CMK were partially rescued by the addition of recombinant BMP2 whose precursor is a candidate for the physiological substrates of PACE4. Further, the transcriptional silencing of PACE4 by its specific siRNAs caused the suppression of both the salivary branching and AQP5 expression in the present organ culture system. These observations strongly support the idea that PACE4 mediates the SMG development.
(キーワード)
Animals / Aquaporin 5 / Bone Morphogenetic Protein 2 / Morphogenesis / Proprotein Convertases / Rats / Salivary Glands / Signal Transduction
Chun Wang, Gang Chen, Junkang Jiang, Lianglin Qiu, Kazuo Hosoi and Chenjuan Yao : Aquaglyceroporins are involved in uptake of arsenite into murine gastrointestinal tissues, The Journal of Medical Investigation : JMI, 56, Suppl, 343-346, Tokushima, Dec. 2009.
(要約)
Aquaglyceroporins (AQGPs) are members of aquaporin (AQP) family and belong to a subgroup of this water channel family; they are transmembrane proteins that transport water as well as glycerol and other solutes of small molecules. Recent studies have also identified that AQGPs are important transporters of trivalent metalloid in some mammalian cells. However, the uptake routes of arsenite in mammals are still less defined. In this study, to understand the routes of arsenite intake in mammals, mice were treated with Hg(II), glycerol, and As(III) and uptake of As(III) into the gastrointestinal tissues was measured. The level of inorganic arsenic (iAs) in gastrointestinal tissues after As(III) stimulation was much higher than Hg(II) +As(III) or glycerol+As(III) group. RT-PCR results showed that AQGPs were extensively expressed in gastrointestinal tissues of mice. We also treated Caco-2 cells with Hg(II) and As(III); the level of iAs in a group treated with Hg(II)+As(III) decreased compared with As(III)-treated group. Our results suggested that AQGPs could be important transporters in arsenite uptake into gastrointestinal tissues of mice, but more data are need to prove if AQGPs is the only pathway involved in As transport in mammals or just one of them.
(キーワード)
Animals / Aquaglyceroporins / Arsenites / Biological Transport / Cell Line / Gastrointestinal Tract / Glycerol / Humans / Male / Mercury / Mice / Mice, Inbred ICR / Signal Transduction
Shingo Kurabuchi, Takanori Matsuoka and Kazuo Hosoi : Hormone-induced granular convoluted tubule-like cells in mouse parotid gland, The Journal of Medical Investigation : JMI, 56, Suppl, 290-295, Tokushima, Dec. 2009.
(要約)
Most striated duct (SD) cells in the adult mouse parotid gland (PAG) have a few small secretory granules. These granules, however, are usually too small and sparse to be detected using light microscopy. Our serial studies have suggested that these PAG SD cells belong to a group of hormone-responsive granular duct cells, similar to the granular convoluted tubule (GCT) cells found in the submandibular gland. These studies also indicate that and some PAG SD cells may be capable of developing a granular cell phenotype under supraphysiological conditions of androgenic and thyroid hormones, leading to more abundant, and more kinds of GCT-specific secretory polypeptides. Here, the cytology of hormone-modulated SD cells, the immunocytochemistry of their secretory products, and their secretory responses to some autonomic agents are reviewed. Finally, the close similarity of the duct systems of the three major salivary glands in mice is critically emphasized.
Kazuo Hosoi, Masataka Murakami and Ivana Novak : The report on the 11th international symposium on exocrine secretion, The Journal of Medical Investigation : JMI, 56, Suppl, 171-178, Tokushima, Dec. 2009.
(キーワード)
Animals / Exocrine Glands / Humans / International Cooperation / Japan
Kazumi Takaishi, Shigemasa Tomioka, Fumiaki Kawano and Kazuo Hosoi : Local anesthetics inhibit bradykinin-stimulated increment of [Ca2+]i in cultured bovine aortic endothelial cells, The Annual Meeting of the American Society of Anesthesiologists, Oct. 2009.
10.
Chenjuan Yao, Ahmad Azlina, Javkhlan Purevjav, Takahiro Hasegawa, Tetsuya Akamatsu and Kazuo Hosoi : Potential down-regulation of parotid gland AQP5 by LPS via cross-coupling of NF-κB/AP1, The 11th International Symposium on Exocrine Secretion, Tokushima 09 ''Exocrine Secretion Mechanism and Disease'', Tokushima, Jul. 2009.
11.
Ahmad Azlina, Nunuk Purwanti, Mileva R. Karabasil, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : Is AQP5 down-regulated via autophagic pathway following chorda tympani denervation?, The International Symposium on Oral Sciences to Improve the Quality of Life, organized by Toshihiko Nagata, Tokushima, Sep. 2008.
12.
Mileva R. Karabasil, Takahiro Hasegawa, Ahmad Azlina, Nunuk Purwanti, Chenjuan Yao, Tetsuya Akamatsu, Shigemasa Tomioka and Kazuo Hosoi : Analyses of rat AQP5 G103D mutant expressed in MDCK-II cells and Xenopus oocytes, The International Symposium on Oral Sciences to Improve the Quality of Life, organized by Toshihiko Nagata, Tokushima, Sep. 2008.
13.
Nunuk Puwanti, Daisuke Tsuji, Mileva Ratko Karabasil, Chenjuan Yao, Takahiro Hasegawa, Tetsuya Akamatsu, Kouji Itou and Kazuo Hosoi : Activation of IL-6/STAT3/Sca-1 system induces proliferation of duct cells in the duct-ligated mouse submandibular gland, The 2nd International Symposium on The Future Direction of Oral Sciences in The 21st Century-Oral Sciences for Our Healthy Life-, organized by Toshihiko Nagata, Tokushima, Dec. 2007.
14.
Mileva R. Karabasil, Kwartarini Murdiastuti, Takahiro Hasegawa, Nunuk Purwanti, Ahmad Azlina, Xuefei Li, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : A naturally occurring point mutation in rat AQP5 influencing its membrane trafficking in and water secretion from salivary gland, The 5th International Conference of Aquaporin, Nara, Jul. 2007.
15.
Xuefei Li, Ahmad Azlina, Mileva R. Karabasil, Nunuk Purwanti, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Effects of autonomic denervation and administration of M3 muscarinic receptor agonist on the AQP5 expression in the rat submandibular gland, The 5th International Conference of Aquaporin, Nara, Jul. 2007.
16.
Chenjuan Yao, Mileva R. Karabasil, Nunuk Purwanti, Ahmad Azlina, Xuefei Li, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : LPS induces down-regulation of AQP5 and AQP1 via Nf-kB and MAPK pathways in the mouse parotid gland, The 5th International Conference of Aquaporin, Nara, Jul. 2007.
17.
Tetsuya Akamatsu, Nunuk Purwanti, Ahmad Azlina, Mileva R. Karabasil, Xuefei Li, Chenjuan Yao, Norio Kanamori and Kazuo Hosoi : Prenatal expression of rat salivary AQP5 is dependent on the submandibular gland development mediated by a subtilisin-like proprotein convertase PACE4, The 5th International Conference of Aquaporin, Nara, Jul. 2007.
18.
Nunuk Puwanti, Daisuke Tsuji, Ahmad Azlina, Mileva Ratko Karabasil, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori, Kouji Itou and Kazuo Hosoi : Alterations of AQP5, cellular markers of duct, and Sca-1 expression in the duct-ligated mouse submandibular gland, The 5th International Conference of Aquaporin, Nara, Jul. 2007.
19.
Mileva R. Karabasil, Takahiro Hasegawa, Nunuk Purwanti, Kwartarini Murdiastuti, Xuefei Li, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Point mutation of AQP5 in Sprague-Dawley rats and its implication for salivary gland physiology, The 1st International Symposium and Workshop on The Future Direction of Oral Sciences in The 21st Century, organized by Bando, E., Awaji, Mar. 2007.
20.
Xuefei Li, Mileva R. Karabasil, Nunuk Purwanti, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Effects of autonomic denervation and administration of muscarinic receptor agonist on the expression of AQPs in the rat submandibular gland, The 1st International Symposium and Workshop on The Future Direction of Oral Sciences in The 21st Century, organized by Bando, E., Awaji, Mar. 2007.
21.
Nunuk Purwanti, Daisuke Tsuji, Mileva Ratko Karabasil, Xuefei Li, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori, Kouji Itou and Kazuo Hosoi : Changes of cellular markers of duct/acinni and side populations in the duct-ligated mouse submandibular gland, The 1st International Symposium and Workshop on The Future Direction of Oral Sciences in The 21st Century, organized by Bando, E., Awaji, Mar. 2007.
22.
Mileva R. Karabasil, Takahiro Hasegawa, Nunuk Purwanti, Kwartarini Murdiastuti, Xuefei Li, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Molecular and cellular analyses of mutant AQP5 which occurred naturally in Spraque - Dawley rats, 3rd International Symposium on Salivary Glands in Honor of Niels Stensen, Okazaki, Japan, Oct. 2006.
23.
Xuefei Li, Mileva R. Karabasil, Nunuk Purwanti, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Effects of autonomic denervation and administration of SNI-2011 on the expression of AQPs in the rat salivary gland, The 3rd International Symposium on Salivary Glands in honor of Niels Stensen, Okazaki, Japan, Oct. 2006.
24.
Chenjuan Yao, Mileva R. Karabasil, Nunuk Purwanti, Xuefei Li, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Tissue kallikrein mK13 is a candidate of the processing enzyme for pro-IL-1β in the mouse submandibular gland, The 3rd International Symposium on Salivary Glands in honor of Niels Stensen, Okazaki, Japan, Oct. 2006.
25.
Tetsuya Akamatsu, Nunuk Purwanti, Mileva R. Karabasil, Xuefei Li, Chenjuan Yao, Norio Kanamori and Kazuo Hosoi : Involvement of a subtilisin-like proprotein convertase, PACE4, in branching morphogenesis and AQP5 expression in the rat embryonic submandibular gland., The 3rd International Symposium on Salivary Glands in honor of Niels Stensen, Okazaki, Japan, Oct. 2006.
26.
Nunuk Purwanti, Daisuke Tsuji, Mileva Ratko Karabasil, Xuefei Li, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori, Kouji Itou and Kazuo Hosoi : The expression of cellular markers of duct/acini and side population dynamics in the duct-ligated mouse submandibular gland., 3rd International Symposium on Salivary Glands in Honor of Niels Stensen, Okazaki, Oct. 2006.
27.
Chenjuan Yao, Mileva R. Karabasil, Nunuk Purwanti, Xuefei Li, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Proteolytic processing of the precursor of interleukin-1β by tissue kallikrein mK13 in the submandibular gland of mice., International symposium on "Medical and Biological Perspectives in Proteases and Their Inhibitors", Satellite meeting of the 20th IUBMB International Congress of Biochemistry and Molecular Biology & the 11th FAOBMB Congress, Awaji, Japan, Jun. 2006.
28.
Tetsuya Akamatsu, Nunuk Purwanti, Mileva R. Karabasil, Xuefei Li, Chenjuan Yao, Norio Kanamori and Kazuo Hosoi : Inhibition of subtilisin-like proprotein convertase PACE4 reduces the branching morphogenesis and expression of AQP5 in the organ culture system of rat embryonic submandibular gland, International symposium on "Medical and Biological Perspectives in Proteases and Their Inhibitors", Satellite meeting of the 20th IUBMB International Congress of Biochemistry and Molecular Biology & the 11th FAOBMB Congress, Awaji, Japan, Jun. 2006.
29.
赤松 徹也, プルワンティ ヌヌク, カラバシル ミレーバ, 李 雪飛, 姚 陳娟, 金森 憲雄, 細井 和雄 : サチライシン様前駆体蛋白質変換酵素PACE4の阻害はラット胎仔顎下腺の分枝形成とAQP5発現を抑制する, 20th IUBMB International Congress of Biochemistry and Molecular Biology & 11th FAOBMB Congress, 第79回日本生化学会大会 & 第29回日本分子生物学会年会, also 第59回日本細胞生物学会大会, 京都, 2006年6月.
30.
Chenjuan Yao, Xuefei Li, Tetsuya Akamatsu, Norio Kanamori and Kazuo Hosoi : Induction, processing, and secretion of IL-1βin the submandibular gland of mice, The 35th International Congress of Physiological Sciences (IUPS2005) and Experimental Biology 2005, San Diego, Mar. 2005.
31.
Tetsuya Akamatsu, Chenjuan Yao, Xuefei Li, Norio Kanamori and Kazuo Hosoi : ECM-bound proprotein convertase PACE4 is involved in the branching morphogenesis of rat submandibular gland, The 35th International Congress of Physiological Sciences (IUPS2005) and Experimental Biology 2005, San Diego, Mar. 2005.
32.
Shingo Kurabuchi and Kazuo Hosoi : Localization and hormonal induction of epidermal growth factor (EGF) and nervegrowth factor (NGF) in the mouse parotid gland, Proceedings of 16th International Congress of the IFAA (Morphological Sciences as the Basis of New Life Sciences in the 21st Century) and 109th Annual Meeting of Japanese Association of Anatomists, 412, Monduzzi Editore, Italy, 2004.
33.
Yoko Yano, Ryoji Ozono, Hidekatu Nakashima, Yoshihiko Oishi, Masayuki Kambe, Kazuo Hosoi and Tetsuya Oshima : Immunohistochemical distributions of the tissue kallikrein-kinin system in ischemic and non-ischemic mouse heart, Journal of Cardiovascular Pharmacology, 42, Suppl.1, S49-S53, Dec. 2003.
(要約)
Kinins have been shown to play a cardioprotective role during myocardial ischemia. However, the localization of each of the components of the kallikrein-kinin system in the heart has not been determined in a cell type-specific manner. Recently, mK1 has been identified as the major tissue kallikrein with the strongest bradykinin-forming activity among the products of the mouse tissue kallikrein gene superfamily. In the study presented here, we investigated the localizations of mK1, kininogen and bradykinin B2 receptors (B2Rs) in ischemic and non-ischemic left ventricles by immunohistochemistry. Kininogen, which contains bradykinin as a surface epitope, was detected by an anti-bradykinin antibody. Changes in the amounts of mK1 and B2R were evaluated by Western blot analysis. Myocardial ischemia was induced by ligation of the left anterior descending coronary artery for 60 min followed by reperfusion for 24 h. mK1 and B2Rs were most abundantly expressed in the vascular endothelium and, to a lesser extent, in fibroblasts. No immunohistochemical signal of these molecules was detected in myocytes. Kininogen was localized in the vascular endothelium and the smooth muscle layer. Myocardial ischemia, although it had no effect on the localization of these molecules, increased the amounts of mK1 and B2R. We have obtained immunohistochemical evidence that all components of the tissue kallikrein-kinin system are present in the mouse heart. The coronary artery is the major site of kallikrein-kinin activity both in ischemic and non-ischemic hearts.
Shingo Kurabuchi, Kazuo Hosoi and Edward W Gresik : Multihormonal regulation of granular convoluted tubular cells expressing mK1, a true tissue kallikrein, in the mouse submandibular gland, Proceedings of the 21st Conference of European Comparative Endocrinologists, 527-530, Monduzzi Editore, Italy, Aug. 2002.
35.
細井 和雄, 松浦 幸子, 倉淵 真悟, 津村 恵子, 多田 淳, 澤 孝賢, 金森 憲雄, 赤松 徹也 : 肥満細胞キニノーゲンと唾液腺カリクレイン-キニン系, The 1st meeting for international collaborative research project 1998-2000., 愛知, 1999年1月.
36.
Kazuo Hosoi, Shingo Kurabuchi, Yamato Kikkawa, Jun Tada, Tetsuya Akamatsu, Naoki Yamanaka, Takuya Matsumoto, Norio Kanamori and Keiko Tsumura : Salivary gland tissue kallikrein family and processing of growth factor precursors and proenzymes., European Journal of Morphology, 36, suppl, 82-85, Cagliari, Italy, Aug. 1998.
Norio Kanamori, Hiroyoshi Sei and Kazuo Hosoi : Phase diagram analysis of sleep-wakefulness cycle: triplet subsystems as the reciprocal control factors., Sleep Res., 24A, 2, Nassau, Bahamas, Sep. 1995.
38.
Takao Ueha, Tadayoshi Kida and Kazuo Hosoi : Relationships between mineralocorticoids and Na+,K+-ATPase in rat submandibular gland, Advances in Physiological Science, 28, 79-84, Budapest, May 1981.
細井 和雄 : Greeting from the director, Facilitating academic partnership with other universities around the world, The University of Tokushima International Center pamphlet, 2009年.