Studies on Water Channel, Aquaporin, Studies on Processing Protease, Studies on Oral Tissue Development (Aquaporin, Water Channel, processing protease, proprotein convertase, Oral Tissue, development)
Book / Paper
Book:
1.
Tetsuya Akamatsu, Chenjuan Yao and Kazuo Hosoi : Methods for animal tissue culture and techniques to prevent cell death, growth defect, and cell mutation, Technical Information Institute Co., Ltd., Apr. 2014.
(Keyword)
salivary gland / tissue culture / induction of differentiation
2.
Kazuo Hosoi, Tetsuya Akamatsu and others : Special Oral Biology - Salivary Glands, Gakken Shoin, Tokyo, Feb. 2006.
3.
Kazuo Hosoi, Tomihiko Higuti, Masanori Kashimata, Rieko Arakaki, Naokatu Arakaki, Jun Tada, Keiko Tsumura, Tetsuya Akamatsu, Kyouko Takeda, Akemichi Ueno, Midori Suenaga, Yoshinobu Baba and others : Experimental Manual for Molecular Cell Biology, 2nd Edition, Nankodo, Tokyo, Apr. 2004.
(Keyword)
In situ hybridization
4.
Hideaki Nagamune, Kazuaki Muramatsu, Masakatsu Higashine, Tetsuya Akamatsu, Yasuhiro Tamai, Yasuo Yonetomi, Akihiko Tsuji, Keisuke Izumi, Takemasa Sakaguchi, Tetsuya Yoshida and Yoshiko Matsuda : Cyclic AMP-inducible procalcitonin processing in thyroidal parafollicular cells is regulated by the Kexin family protease, PC1., IOS Press, Amsterdam, Mar. 1997.
(Summary)
カルシウム調節ホルモンであるカルシトニンは甲状腺傍濾胞細胞より分泌されるが, 不活性な前駆体として合成された後, プロセシングを受けて初めて活性化される. 本研究ではcAMP刺激により著増するカルシトニンのプロセシングがKexin family proteaseに属する酵素PC1により調節されることを初めて明らかにした.
(Keyword)
Calcitonin / Processing / PC1
Academic Paper (Judged Full Paper):
1.
Hiroshi Yoshimura, Sugai Tokio, Kato Nobuo, Tominaga Takashi, Tominaga Yoko, Takahiro Hasegawa, Chenjuan Yao and Tetsuya Akamatsu : Interplay between non-NMDA and NMDA receptor activation during oscillatory wave propagation: Analyses of caffeine-induced oscillations in the visual cortex of rats, Neural Networks, Vol.79, 141-149, 2016.
(Summary)
Generation and propagation of oscillatory activities in cortical networks are important features of the brain. However, many issues related to oscillatory phenomena are unclear. We previously reported neocortical oscillation following caffeine treatment of rat brain slices. Input to the primary visual cortex (Oc1) generates N-methyl-d-aspartate (NMDA) receptor-dependent oscillations, and we proposed that the oscillatory signals originate in the secondary visual cortex (Oc2). Because non-NMDA and NMDA receptors cooperate in synaptic transmission, non-NMDA receptors may also play an important role in oscillatory activities. Here we investigated how non-NMDA receptor activities contribute to NMDA receptor-dependent oscillations by using optical recording methods. After induction of stable oscillations with caffeine application, blockade of NMDA receptors abolished the late stable oscillatory phase, but elicited 'hidden' non-NMDA receptor-dependent oscillation during the early depolarizing phase. An interesting finding is that the origin of the non-NMDA receptor-dependent oscillation moved from the Oc1, during the early phase, toward the origin of the NMDA receptor-dependent oscillation that is fixed in the Oc2. In addition, the frequency of the non-NMDA receptor-dependent oscillation was higher than that of the NMDA receptor-dependent oscillation. Thus, in one course of spatiotemporal oscillatory activities, the relative balance in receptor activities between non-NMDA and NMDA receptors gradually changes, and this may be due to the different kinetics of the two receptor types. These results suggest that interplay between the two receptor types in the areas of Oc1 and Oc2 may play an important role in oscillatory signal communication.
Hiroshi Yoshimura, 川邊 真道, 須貝 外喜夫, 加藤 伸郎, Takahiro Hasegawa, Chenjuan Yao and Tetsuya Akamatsu : Influences of oral impairment on neural oscilltion and wave propagation in the neocortex of rats, Neuroscience Research Suppl, Vol.Suppl, 2015.
3.
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, Vol.306, No.1, E100-E108, 2014.
(Summary)
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.
Hiroshi Yoshimura, Tokio Sugai, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Nobuo Kato : Age-dependent emergence of caffeine-assisted voltage oscillations in the endopiriform nucleus of rats., Neuroscience Research, Vol.76, No.1-2, 16-21, 2013.
(Summary)
The gustatory insular cortex (IC) is connected with not only the somatosensory cortex, but also the endopiriform nucleus (EPN). We have previously revealed that low-frequency electrical stimulation to the IC can elicit membrane potential oscillations at a frequency of 8-10 Hz in the somatosensory cortex of rat brain slices under bath-application of caffeine. Using the same procedure, we investigated whether the EPN has the ability to generate oscillations, and whether such oscillations emerge age-dependently. Electrical stimulations were delivered to the IC, and field potentials were recorded from the EPN. In the case of slices made from mature rats, stable field potential oscillations at 8-10 Hz were induced in the EPN after repetitive stimulations. Optical recordings revealed that signals traveled from the IC to the EPN by way of the claustrum. Generation of oscillations was N-methyl-d-aspartate (NMDA) receptor activity-dependent, since oscillatory phases disappeared following application of NMDA receptor antagonist. In slices from immature rats, however, oscillations were not induced. IC stimulation can thus age-dependently elicit membrane potential oscillations in the EPN, and the EPN oscillations were NMDA receptor activity-dependent. These findings suggest that developmental changes in properties of the EPN might contribute to development of information integration, including gustatory information.
(Keyword)
aging / Animals / Caffeine / Central Nervous System Stimulants / Cerebral Cortex / Electric Stimulation / Membrane Potentials / Organ Culture Techniques / Rats / Rats, Wistar
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, Vol.301, No.5, G814-G824, 2011.
(Summary)
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, Vol.34, No.6, 668-680, 2011.
(Summary)
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, Vol.301, No.3, C667-C678, 2011.
(Summary)
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, Vol.40, No.8, 651-658, 2011.
(Summary)
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.
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, Vol.103, No.2, 69-86, 2011.
(Summary)
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.
(Keyword)
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, Vol.299, No.5, G1106-G1117, 2010.
(Summary)
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, Vol.177, No.2, 724-734, 2010.
(Summary)
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, Vol.56, No.1, 2, 55-63, 2009.
(Summary)
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.
(Keyword)
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
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, Vol.325, No.2, 434-443, 2009.
(Summary)
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.
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, Vol.295, No.1, G112-G123, 2008.
(Summary)
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, Vol.236, No.1, 314-320, 2007.
(Summary)
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, Vol.291, No.6, GI1081-GI1088, 2006.
(Summary)
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.
(Keyword)
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, Vol.1763, No.4, 337-344, 2006.
(Summary)
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.
(Keyword)
Animals / Aquaporin 5 / Cell Line / Cell Membrane / Cyclic AMP-Dependent Protein Kinases / Dogs / Green Fluorescent Proteins / Protein Transport / Recombinant Fusion Proteins
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, Vol.281, No.12, 7968-7976, 2006.
(Summary)
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.
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, Vol.290, No.2, F478-F485, 2006.
(Summary)
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, Vol.116, No.2, 213-222, 2005.
(Summary)
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.
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, Vol.288, No.6, G1283-G1291, 2005.
(Summary)
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.
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, Vol.446, No.6, 641-651, 2003.
(Summary)
The expression and localization of aquaporins (AQP1-AQP5), members of the water channel family, in the developing rat submandibular gland were analysed using RT-PCR, Northern blotting and immunohistochemistry to explore their relation to the development of this salivary gland. RT-PCR analysis revealed unique expression patterns of each AQP. AQP1 was expressed constitutively during prenatal development, whereas the expression of AQP5 became more intense in the course of development from embryonic day 16.5 (E16) to E20. These expression patterns concurred with the results of Northern blot analysis. AQP3 and AQP4 mRNAs in the prenatal development were not detected in Northern blots, although they were detected by RT-PCR. During postnatal development, AQP5 and AQP1 mRNAs were expressed continuously, but no message for AQP3 or AQP4 was detected. AQP2 mRNA was not detected during either prenatal or postnatal development in this tissue. Immunohistochemical studies revealed that AQP5 was first localized at the apical membrane of proacinar cells at E18, and then became clearly distributed at the apical membrane of acinar cells in accordance with the differentiation and establishment of the mature acini. In addition, some vasculature also showed immunoreactivity for AQP5. AQP1 was immunolocalized in the blood vessels, including capillaries, of the gland throughout development. These observations suggest the existence of transcriptional regulation of rat AQP5, which is one of the most probable regulators of saliva production and secretion, during the establishment of the functional submandibular salivary gland.
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, Vol.445, No.3, 405-412, 2002.
Kwartarini Murdiastuti, Osamu Miki, Chenjuan Yao, Nahid Most. Parvin, Tetsuya Akamatsu, Tetsuya Akamatsu, Norio Kanamori, Norio Kanamori and Keiko Tsumura : 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, Vol.445, No.3, 405-412, 2002.
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, Vol.1542, No.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), Vol.3, No.1, 75-77, 2002.
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, Vol.69, No.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), Vol.12, No.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), Vol.2, No.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, Vol.272, No.2, 410-415, 2000.
(Summary)
Expressions of mRNAs for four subtilisin-like proprotein convertases (SPCs: furin, PACE4, PC6, and PC8) and bone morphogenetic protein 4 (BMP4) in the rat molar tooth during development were analyzed by Northern blotting, reverse transcriptase-polymerase chain reaction (RT-PCR), and in situ hybridization to explore the possible involvement of SPCs in the processing of proBMPs. We found a temporospacial expression of PACE4, but not one of the other SPCs, in this tissue; i.e., RT-PCR analysis revealed that the level of PACE4 mRNA, but not that of the other SPC mRNAs became high around the second postnatal day. This increase was in good accordance with the increase in BMP4 mRNA, indicating an apparent association of these molecules with the differentiation and establishment of functional ameloblasts and odontoblasts. During dentinogenesis, PACE4 mRNA was localized in the ameloblasts and odontoblasts. These observations suggest that PACE4 plays a crucial role in dentinogenesis, especially via the activation of BMPs.
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, Vol.101, No.4, 531-540, 2000.
(Summary)
The connective tissue-type mast cells present in the submandibular gland (SMG) and peritoneal cavity of rats were found to express kininogens (KGs), the expression of which was demonstrated by Western blotting, reverse transcription-polymerase chain reaction (RT-PCR), RT-PCR Southern blotting, and light- and electron-microscopic immunocytochemistry. In the SMG, the analysis of cDNA amplified by RT-PCR revealed that the molecular species of mRNAs expressed were high-molecular-weight (HMW)-K KG and T-I KG. Light microscopic immunocytochemistry exclusively localized the KG protein(s) in the mast cells present in the SMG. The signals in the mast cells were very strong, but no positive reaction was observed in the granular convoluted tubular cells, acinar cells or striated duct cells. As determined by using electron microscopy, extremely strong labelling with immunogold was observed in the secretory granules of the mast cells, but no labelling in their nucleus or cytoplasm. Analysis by Western blotting and RT-PCR Southern blotting indicated that both protein and mRNA of KGs were present in the mast cells separated from the peritoneal cavity, indicating de novo synthesis of KG in these cells. Preliminary experiments implied that the connective tissue-type mast cells in other rat tissues also expressed KG.
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, Vol.266, No.2, 443-447, 1999.
Aquaporin 5 / Water channel / Trafficking / Cytoskeleton
33.
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, Vol.216, No.4/5, 481-488, 1999.
Processing protease / PACE4 / Gene structure / Alternative splicing
35.
Tetsuya Akamatsu, Shigeo Daikoku, Hideaki Nagamune, Shigeru Yoshida, Kenji Mori, Akihiko Tsuji and Yoshiko Matsuda : Developmental expression of a novel Kexin family protease, PACE4E, in the rat olfactory system., Histochemistry and Cell Biology, Vol.108, No.2, 95-103, 1997.
(Summary)
本研究では新規Kexin family proteaseであるPACE4Eがラット脳発生過程において時期·空間特異的に発現することを明らかにし, 特に嗅球においては僧帽細胞に極めて特異的に発現し, 僧帽細胞の分化成熟と密接に関わることを見い出した.
(Keyword)
Processing protease / Olfactory system / Mitral cell
36.
Kenji Mori, Sachiko Kii, Akihiko Tsuji, Masami Nagahama, Akiyoshi Imamaki, Keiko Hayashi, Tetsuya Akamatsu, Hideaki Nagamune and Yoshiko Matsuda : A Novel Human PACE4 Isoform, PACE4E Is an Active Processing Protease Containing a Hydrophobic Cluster at the Carboxy Terminus, The Journal of Biochemistry, Vol.121, No.5, 941-948, 1997.
Hideaki Nagamune, Kazuaki Muramatsu, Tetsuya Akamatsu, Yasuhiro Tamai, Keisuke Izumi, Akihiko Tsuji and Yoshiko Matsuda : Distribution of the kexin family proteases in pancreatic islets., --- PACE4C is specifically expressed in B cells of pancreatic islets ---, Endocrinology, Vol.136, No.1, 357-360, 1995.
(Summary)
Kexin Family Proteasesはペプチドホルモン等のプロセシング酵素として同定され, 血糖調節ホルモンのインスリン等のプロセシングにも関与することが示唆されていたが, 本研究では我々が新規にクローニングしたPACE4CをはじめとするKexin Family Proteasesとインスリン等との膵島細胞での局在, 共存関係を初めて明らかにし, その成果は雑誌表紙にも掲載された.
Akihiko Tsuji, Tetsuya Akamatsu, Hideaki Nagamune and Yoshiko Matsuda : Identification of targeting proteinase for rat α1-macroglobulin in vivo., --- Mast-cell tryptase is a major component of the α1-macroglobulin-proteinase complex endocytosed into rat liver lysosomes. ---, The Biochemical Journal, Vol.298, No.1, 79-85, 1994.
Chenjuan Yao, Tetsuya Akamatsu, Takahiro Hasegawa and Hiroshi Yoshimura : The Defense System of Oral Cavity: Lipopolysaccharide Induced Inflammatory Response in the Mice Salivary Gland, BIT's 1st Annual World Congress of Oral & Dental Medicine Conference Abstract Book, 107, 2014.
Tetsuya Akamatsu, Chenjuan Yao, Takahiro Hasegawa and Hiroshi Yoshimura : Salivary Gland Development, Differentiation, and Regeneration - Role of Subtilisin-like Proprotein Convertase PACE4/SPC4, BIT's 1st Annual World Congress of Oral & Dental Medicine Conference Abstract Book, 59, 2014.
Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Hiroshi Yoshimura : Enhanced ubiquitylation and downregulation of aquaporin-5 by ubiquitin ligases, Journal of Oral Biosciences, Vol.Suppl., 204, 2014.
(Keyword)
water channel / ubiquitylation / downregulation / AQP5
4.
Chenjuan Yao, Tetsuya Akamatsu, Takahiro Hasegawa and Hiroshi Yoshimura : Expression of water channel AQP5 in regeneration model of salivary gland, Journal of Oral Biosciences, Vol.Suppl., 204, 2014.
(Keyword)
salivary gland / regeneration / water channel / AQP5
5.
Minegishi Makoto, Tetsuya Akamatsu, Chenjuan Yao, Takahiro Hasegawa and Hiroshi Yoshimura : Induced expression of subtilisin-like proprotein convertase in regeneration model of salivary gland, Journal of Oral Biosciences, Vol.Suppl., 138, 2014.
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, Vol.21, No.4, 1182, Feb. 2020.
Tetsuya Akamatsu : Structure and Function of the Salivary Gland, Journal of Oral Health and Biosciences, Vol.28, No.2, 77-86, Feb. 2016.
(Summary)
The salivary gland is developed under the epithelial-mesenchymal interaction and formed by repeating branching morphogenesis, which is a common process in the development of the glandular tissues. Although the differentiation and maturation of salivary gland is continued into early postnatal life, the fundamental ability of saliva secretion is already expressed after birth.These development, differentiation, and maturation of salivary gland are regulated by many growth/ differentiation factors, which are initially synthesized as inactive precursors and activated by the limited proteolysis. On the other hand, saliva secretion is one of the important physiological functions of the salivary gland and occurs dependently on the increase of the osmolality in the lumen through two pathways, paracellular and transcellular pathways. It is revealed that a water channel aquaporin 5 (AQP5) is involved in saliva secretion through the latter pathway. Saliva contains various components, which express various physiological functions of saliva. Because the oral cavity is confronted with hazards of various allergens from the outside continuously, the salivary gland produces and secretes various factors as the defense system. Saliva is, therefore, one of an important body fluid to maintain the oral health, and the decrease of saliva secretion causes xerostomia/dry mouth, which affects not only oral disease and dysfunction but also systemic disease. We previously reported the involvement of a subtilisin-like proprotein convertase PACE4 in the development and differentiation of salivary gland, lipopolysaccharide-mediated induction of inflammatory cytokines in the salivary gland, degradation of salivary AQP5 by parasympathetic denervation, and so on. This review will describe the structure and function of the salivary gland, from its development to functional expression and regulation.
● CiNii @ National Institute of Informatics (NAID): 120005823527
(Tokushima University Institutional Repository: 109872, CiNii: 120005823527)
3.
Tetsuya Akamatsu : Physiological role of a subtilisin-like proprotein convertase, PACE4, in submandibular gland development, Journal of Oral Biosciences, Vol.52, No.2, 81-93, May 2010.
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, Vol.37, 26-31, Mar. 2001.
5.
Yoshiko Matsuda, Akihiko Tsuji, Hideaki Nagamune, Tetsuya Akamatsu, Chiemi Hine, Kazuaki Muramatsu, Kenji Mori, Yasuhiro Tamai and Kayoko Yonemoto : Novel members of mammalian Kexin family proteases, PACE4C, PACE4D, PC7A and PC7B., Advances in Experimental Medicine and Biology, Vol.389, 63-71, Sep. 1996.
(Summary)
ペプチドホルモンや増殖因子等は不活性な前駆体として合成され, 特定の塩基性アミノ酸部位での限定切断を経て初めて活性化される. Kexin family proteaseはこのプロセシングを担う重要な酵素であり, 本研究では我々が新規メンバーをクローニングし, その構造, 組織分布等を明らかにした.
(Keyword)
Processing protease
Proceeding of International Conference:
1.
Chenjuan Yao, Tetsuya Akamatsu, Takahiro Hasegawa and Hiroshi Yoshimura : Induced expression of a subtilisin-like proprotein convertase PACE4 in the regeneration model of rat submandibular gland., The 4th International Symposium on Salivary Glands in Honor of Niels Stensen, Okazaki (Japan), Nov. 2016.
2.
Tetsuya Akamatsu, Chenjuan Yao, Takahiro Hasegawa and Hiroshi Yoshimura : Sexual difference in the regeneration model of the rat submandibular gland., The 4th International Symposium on Salivary Glands in Honor of Niels Stensen, Okazaki (Japan), Nov. 2016.
3.
Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Hiroshi Yoshimura : Post-translational modifications of water channel aquaporin-5 in salivary gland cells, Oral Neuroscience 2016, Oral Neuroscience 2016, Osaka, Oct. 2016.
4.
Hiroshi Yoshimura, Tetsuya Akamatsu, Chenjuan Yao and Takahiro Hasegawa : Synaptic plasticity in the brain -Roles of NMDA receptor- (Invited lecture at Nantong University), Sep. 2016.
(Keyword)
synaptic plasticity / NMDA-receptors
5.
Chenjuan Yao, Tetsuya Akamatsu, Takahiro Hasegawa and Hiroshi Yoshimura : The Defense System of Oral Cavity: Lipopolysaccharide Induced Inflammatory Response in the Mice Salivary Gland, Haikou, China, Nov. 2014.
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, Vol.56, No.Suppl, 398-400, Tokushima, Dec. 2009.
(Summary)
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, Vol.56, No.Suppl, 287-289, Tokushima, Dec. 2009.
(Summary)
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, Vol.56, No.Suppl, 273-276, Tokushima, Dec. 2009.
(Summary)
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, Vol.56, No.Suppl, 253-254, Tokushima, Dec. 2009.
(Summary)
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, Vol.56, No.Suppl, 241-246, Tokushima, Dec. 2009.
(Summary)
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.
(Keyword)
Animals / Aquaporin 5 / Bone Morphogenetic Protein 2 / Morphogenesis / Proprotein Convertases / Rats / Salivary Glands / Signal Transduction
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, Vol.36, No.suppl, 82-85, Cagliari, Italy, Aug. 1998.
Hideaki Nagamune, Kazuaki Muramatsu, Masakatsu Higashine, Tetsuya Akamatsu, Yasuhiro Tamai, Yasuo Yonetomi, Akihiko Tsuji, Keisuke Izumi, Takemasa Sakaguchi, Tetsuya Yoshida and Yoshiko Matsuda : Cyclic AMP-inducible procalcitonin processing in thyroidal parafollicular cells is regulated by the Kexin family protease, PC1., International Symposium on Medical Aspects of Protease Inhibitors; In "Medical Aspects of Proteases and Protease Inhibitors": Biomedical and Health Research, Vol.15, 22-33, Amsterdam, May 1997.
Maeda Saori, Hiroshi Yoshimura, Miyaji Yuji, Hiroyuki Kanayama, Takahiro Hasegawa, Chenjuan Yao and Tetsuya Akamatsu : Increase in theta-band EEG activities under tasting chocolate with unmatched odor stimulation, 第41回日本神経科学大会, Jul. 2018.
(Keyword)
electroencephalogram / theta wave
5.
前田 さおり, Hiroshi Yoshimura, 宮地 ゆうじ, 金山 宏幸, Takahiro Hasegawa, Chenjuan Yao and Tetsuya Akamatsu : Theta band brain activity during checking the unmatched olfactory-taste information, 第95回日本生理学大会, Mar. 2018.
前田 さおり, Hiroshi Yoshimura, 宮地 ゆうじ, Takahiro Hasegawa, Chenjuan Yao and Tetsuya Akamatsu : Influences of Match/Mismatch of taste and odor on taste perception: An electroencephalogram frequency analysis study, 日本味と匂学会第51回大会, Sep. 2017.
Maeda Saori, Miyachi Yuki, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Hiroshi Yoshimura : Influences of olfactory stimulation on taste perception: An electroencephalogram frequency analysis study, The 94th Annual Meeting of the Physiological Science of Japan, Mar. 2017.
Tetsuya Akamatsu, Chenjuan Yao, Takahiro Hasegawa and Hiroshi Yoshimura : Sexual difference in regeneration of salivary gland, 第58回歯科基礎医学会学術大会, Aug. 2016.
12.
Shimatani Tatsuya, Minegishi Makoto, Tetsuya Akamatsu, Chenjuan Yao, Takahiro Hasegawa and Hiroshi Yoshimura : Induction of a subtilisin-like proprotein convertase PACE4 in regeneration model of salivary gland Part II, 第58回歯科基礎医学会学術大会, Aug. 2016.
13.
Hiroshi Yoshimura, 川邊 真道, 須貝 外喜夫, 加藤 伸郎, Takahiro Hasegawa, Chenjuan Yao and Tetsuya Akamatsu : Influences of oral impairment on neural oscilltion and wave propagation in the neocortex of rats, 第38回日本神経科学大会, Jul. 2015.
14.
Tetsuya Akamatsu : Structure and Function of the Salivary Gland, Jun. 2015.
15.
Chenjuan Yao, Tetsuya Akamatsu, Takahiro Hasegawa and Hiroshi Yoshimura : Expression of water channel AQP5 in regeneration model of salivary gland, 第56回歯科基礎医学会, Sep. 2014.
(Keyword)
salivary gland / regeneration / water channel / AQP5
16.
Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Hiroshi Yoshimura : Enhanced ubiquitylation and downregulation of aquaporin-5 by ubiquitin ligases, 第56回歯科基礎医学会, Sep. 2014.
(Keyword)
water channel / ubiquitylation / downregulation / AQP5
17.
Minegishi Makoto, Tetsuya Akamatsu, Chenjuan Yao, Takahiro Hasegawa and Hiroshi Yoshimura : Induced expression of subtilisin-like proprotein convertase in regeneration model of salivary gland, 第56回歯科基礎医学会(福岡国際会議場/福岡県), Sep. 2014.
Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Hiroshi Yoshimura : ヒト唾液腺HSG細胞における構成的なAQP5の取り込み, Journal of Oral Biosciences, Vol.55, No.Suppl., 199, 2013.
22.
Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : 唾液腺細胞におけるアクアポリン5の翻訳後修飾, Journal of Oral Biosciences, Vol.53, No.Supplement, 104, Sep. 2011.
23.
Purevjav Javkhlan, Yuka Hiroshima, Ahmad Azlina, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu, Toshihiko Nagata and Kazuo Hosoi : Calprotectin expression in the mouse salivary gland: Induction by lipopolysaccharide, 第52回歯科基礎医学会(2010年9月20-22日), Sep. 2010.
24.
Yuka Hiroshima, Purevjav Javkhlan, Ahmad Azlina, Takahiro Hasegawa, Tetsuya Akamatsu and Kazuo Hosoi : Porphyromonas gingivalis由来LPSはヒト好中球からのレジスチンの遊離を促進する, 第52回歯科基礎医学会(2010年9月20-22日), Sep. 2010.
25.
Takahiro Hasegawa, Ahmad Azlina, Purevjav Javkhlan, Yuka Hiroshima, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : 唾液腺細胞におけるアクアポリン5リン酸化のシグナル伝達機構, 第52回歯科基礎医学会(2010年9月20-22日), Sep. 2010.
26.
Purevjav Javkhlan, Yuka Hiroshima, Ahmad Azlina, Takahiro Hasegawa, Chenjuan Yao, Tetsuya Akamatsu, Toshihiko Nagata and Kazuo Hosoi : Calprotectin expression in the mouse salivary gland: Induction by lipopolysaccharide, Journal of Oral Biosciences, Vol.52, No.supplement, 94, Sep. 2010.
27.
Yuka Hiroshima, Javkhlan Purevjav, Ahmad Azlina, Takahiro Hasegawa, Tetsuya Akamatsu and Kazuo Hosoi : Porphyromonas gingivalis由来LPSはヒト好中球からのレジスチンの遊離を促進する, Journal of Oral Biosciences, Vol.52, No.supplement, 148, Sep. 2010.
28.
Takahiro Hasegawa, Ahmad Azlina, Javkhlan Purevjav, Yuka Hiroshima, Chenjuan Yao, Tetsuya Akamatsu and Kazuo Hosoi : 唾液腺細胞におけるアクアポリン5リン酸化のシグナル伝達機構, Journal of Oral Biosciences, Vol.52, No.supplement, 164, Sep. 2010.
29.
プルワンティ ヌヌク, Daisuke Tsuji, アズリナ アハマド, カラバシル ミレーバ, Chenjuan Yao, Takahiro Hasegawa, Tetsuya Akamatsu, Kouji Itou and Kazuo Hosoi : Transient increase in IL-6 by duct ligation leading to continuous elevation of Sca-1, a stem cells marker, in the mouse submandibular gland, 第49回歯科基礎医学会学術大会, Aug. 2007.
30.
プルワンティ ヌヌク, Daisuke Tsuji, カラバシル ミレーバ, Xuefei Li, Chenjuan Yao, Tetsuya Akamatsu, Norio Kanamori, Kouji Itou and Kazuo Hosoi : Effect of duct ligation on the expression of cellular markers of duct/acini and side population dynamics of the mouse submandibular gland, 第48回歯科基礎医学会学術大会, Sep. 2006.
Et cetera, Workshop:
1.
Chenjuan Yao, Takahiro Hasegawa, Tetsuya Akamatsu and Hiroshi Yoshimura : マウス耳下腺における水チャネルAQP5のイソプロテレノールによるdouwn-regulationの機構, Journal of Oral Biosciences, Vol.54, No.Suppl., 153,
2.
Shimatani Tatsuya, Minegishi Makoto, Tetsuya Akamatsu, Chenjuan Yao, Takahiro Hasegawa and Hiroshi Yoshimura : Induction of a subtilisin-like proprotein convertase PACE4 in regeneration model of salivary gland Part II, Journal of Oral Biosciences, Vol.Suppl., 415, Aug. 2016.
3.
Tetsuya Akamatsu, Chenjuan Yao, Takahiro Hasegawa and Hiroshi Yoshimura : Sexual difference in regeneration of salivary gland, Journal of Oral Biosciences, Vol.Suppl., 451, Aug. 2016.
4.
Tetsuya Akamatsu : 基礎系教育講演, Journal of Oral Health and Biosciences, Vol.28, No.2, 97, Feb. 2016.
5.
Tetsuya Akamatsu, Chenjuan Yao, Takahiro Hasegawa and Hiroshi Yoshimura : 唾液腺再生への subtilisin-like proprotein convertase PACE4の関与, 自然科学研究機構生理学研究所研究会「唾液腺形態形成研究会~機能解析から器官再生へ~」, Aug. 2014.
Hiroshi Yoshimura, Takahiro Hasegawa, Chenjuan Yao and Tetsuya Akamatsu : 細胞内cAMP上昇が大脳皮質味覚野から口腔体性感覚野への信号伝播速度に与える影響,第54回歯科基礎医学会(郡山)2012年9月14-16日, Sep. 2012.
8.
Chenjuan Yao, Takahiro Hasegawa, Tetsuya Akamatsu and Hiroshi Yoshimura : マウス耳下腺における水チャネルAQP5のイソプロテレノールによるdouwn-regulationの機構, Journal of Oral Biosciences, Vol.54, No.Suppl., 153, Sep. 2012.
9.
Hiroshi Yoshimura, Takahiro Hasegawa, Chenjuan Yao and Tetsuya Akamatsu : 細胞内cAMP上昇が大脳皮質味覚野から口腔体性感覚野への信号伝播速度に与える影響, Journal of Oral Biosciences, Vol.54, No.Suppl., 131, Sep. 2012.
Report:
1.
Kazuo Hosoi, Norio Kanamori, Tetsuya Akamatsu and Chenjuan Yao : Molecular mechanism of expression and regulation of water channel, esp. aquaporins in the exocrine gland, , Report of research project, grant in-aid for scientific research (B), Tokushima, Mar. 2009.
2.
Kazuo Hosoi, Norio Kanamori, Tetsuya Akamatsu and Chisato Kosugi : Induction of inflammation cytokine and defense system of oral cavity: a trial in regeneration medicine, Report of research project, Grant-in-Aid for Exploratory Research, Tokushima, Mar. 2006, Tokushima, Mar. 2006.
3.
Kazuo Hosoi, Nobuyoshi Nakajo, Tatsuji Haneji and Tetsuya Akamatsu : Inflammation by mast cell kininogen and its suppression Development of a new drug, , Report of research project, grant in-aid for scientific research (B)(2), Tokushima, Mar. 2004.
4.
Kazuo Hosoi, Norio Kanamori, Tetsuya Akamatsu, Jun Tada and Keiko Tsumura : : Expression and regulation of a water channel, aquaporin, in the exocrine gland, Report of research project, grant in-aid for scientific research (C)(2), Tokushima, Mar. 2003.
5.
Kazuo Hosoi, Norio Kanamori, Sachiko Matsuura, Tetsuya Akamatsu and Takamasa Sawa : Molecular mechanism of expression and regulation of water channel proteins aquaporins in the salivary gland cells, Report of research project, grant in-aid for scientific research (C)(2), Tokushima, Mar. 2001.
Development of low invasive long-term imaging technique for studying salivary gland Ca2+ response and functional regeneration (Project/Area Number: 16K11484 )
Elucidation of Molecular Mechanism of the Salivary Gland Regeneration ~ Toward the application to the Regenerative Medicine for Xerostomia ~ (Project/Area Number: 26462814 )
Elucidation of the molecular mechanism on the differentiation/maturation and functional expression of salivary acinar cell toward conquest of xerostomia. (Project/Area Number: 23592737 )
Molecular mechanisms of expression and regulation of function of water channel proteins aquaporins in the salivary gland cells (Project/Area Number: 11671844 )