1998 Fiscal Year Annual Research Report

体液・循環調節に関与するペプチドホルモン受容体に関する比較分子生物学的研究(塩類細胞の分子解剖と分化誘導機構の解明)

Research Project

Project/Area Number	09102008
Research Institution	Tokyo Institute of Technology
Principal Investigator	広瀬茂久東京工業大学, 生命理工学部, 教授 (10134199)
Co-Investigator(Kenkyū-buntansha)	萩原啓実東京工業大学, 生物実験センター, 助教授 (90189465) 竹井祥郎東京大学, 海洋研究所, 教授 (10129249)
Keywords	循環調節 / ANP受容体 / 海水適応 / 塩類細胞 / ウナギ / 広塩性魚類 / 酸性適応 / ウグイ
Research Abstract	循環系の恒常性維持機構をよりよく理解するために,比較分子生物学的なアプローチを試み,以下の成果を得た。1)ウナギの海水適応に重要な分子として同定したKチャネルが塩類細胞の微細管系に局在することを免疫電子顕微鏡法で明らかにすると共に,この研究を哺乳類にまで拡げ,ラットから新型Kチャネルをクローニングすることに成功した。2)pH3.5という硫酸酸性湖である恐山湖にすむウグイを材料に酸性適応に重要な分子の一つとして,V-ATPaseをウグイのエラよりクローニングし,ノーザン分析や抗体を用いるウエスタン分析により,酸性下ではその発現が塩類細胞で昂進することを明らかにした。このほかにも,イオン輸送や水輸送に重要なトランスポーターやチャネルも候補分子として同定している。3)血圧に影響を与えない微量濃度で心房性ナトリウム利尿ペプチド(ANP)がウナギの飲水の抑制と血漿アンギオテンシン濃度の低下を引き起こすという興味深い現象を見いだした。さらにANPによる飲水の抑制が,飲水誘起ホルモンであるアンギオテンシンの低下を介する間接作用ではなく,直接作用であることも明らかにした。4)塩類細胞は,魚類において淡水中での塩の摂取や海水中でのNaClの排出に主要な役割を果たしているが,Clチャンネルの局在,すなわち外界側にあるか血液側にあるかにより輸送の方向が決められる。ティラピアは淡水と海水双方によく適応する広塩性魚であるが,ティラピアの鰓のcDNAライブラリーから2種のCLC型のClチャンネルをクローン化した。そのうち,補乳類のCLC-5に相同なチャンネルは浸透圧調節器官である鰓,腸,腎臓などに多く発現しており,鰓では塩類細胞に局在して発現していることを明らかにした。現在,電顕レベルの免疫細胞化学法を用いて,塩類細胞における局在部位を明らかにしつつある。

Research Products

(23 results)

All Other

All Publications (23 results)

[Publications] Suzuki,Y.et al.: "Identification by differential display of a hypertonicity-inducible inward rectifier pottasium channel highly expressed in chloride cells." J.Biol.Chem.274(in press). (1999)
[Publications] Hagiwara,H.et al.: "Deceleration by angiotensin II of the differentiation and bone formation of rat calvarial osteoblastic cells." J.Endocrinol.156. 543-550 (1998)
[Publications] Hiruma,Y.et al.: "Endothelins inhibit the mineralization of osteoblastic MC3T3-E1 cells through thr A-type endothelin receptor." Am.J.Physiol.275. R1099-R1105 (1998)
[Publications] Kashiwagi,M.et al.: "Cloning,properties and tissue distribution of natriuretic peptide receptor-A of euryhaline eel,Anguilla japonica." Eur.J.Biochem.259. 204-211 (1999)
[Publications] Nagase,M.et al.: "Genomic organization and regulation of expression of the lectin-like oxidized low-density lipoprotein receptor(LOX-1)gene." J.Biol.Chem.273. 33702-33707 (1998)
[Publications] Tsuchida,T.et al.: "Effects of homologous atrial natriuretic peptide on drinking and plasma angiotensin II level in eels." Am.J.Physiol.230. R1605-R1610 (1998)
[Publications] Miyazaki,H.et al.: "Molecular cloning of CLC channels in Oreochromis mossambicus and their functional complementation of yeast CLC gene mutant" Biochem.Biophys.Res.Commun.(in press). (1999)
[Publications] Takei,Y.et al.: "Tetrapod-type[Asp1]angiotensin is present in a primitive holostean fish,Amia calva." Gen.Comp.Endocrinol.110. 140-146 (1998)
[Publications] Sakaguchi,H.et al.: "Characterisation of C-type natriuretic peptide receptors in the gill of dogfish Triakis scyllia." J.Endocrinol.156. 127-134 (1998)
[Publications] Tierney,M.L.et al.: "A radioimmunoassay for the detection of angiotensin II in elasmobranch fish." Gen.Comp.Endocrinol.111. 299-305 (1998)
[Publications] Takei,Y.et al.: "Antidiuretic effect of eel ANP infused at physiological doses in seawater-adapted eels,Anguilla japonica." Zool.Sci.15. 399-404 (1998)
[Publications] Suzuki,N.et al.: "Amino acid sequences of N-terminal procalcitonin of some vertebrates." Nether.J.Zool.48. 349-360 (1998)
[Publications] Hamano,K.et al.: "Direct vasoconstrictor action of homologous angiotensin II on isolated aterial ring preparation in an elasmobranch fish." J.Endocrinol.158. 419-423 (1997)
[Publications] McKendry,J.E.et al.: "The effects of netriuretic peptides on catecholamine release in American eel(Anguilla rostrata),rainbow trout(Oncorhynchus mykiss),and spiny dogfish(Squalus acanthias)." Fish Physiol.Biochem.(in press). (1999)
[Publications] Takei,Y.: "Comparative endocrinology of natriuretic peptide systems." Int.Rev.Cytol.(in press). (1999)
[Publications] Hazon,N.et al.: "he renin-angiotensin system in elasmobranch fish : A review." J.Exp.Zool.(in press). (1999)
[Publications] Takei,Y.: "Comparative physiology of body fluid regulation. in eels." Jpn.J.Physiol.(in press). (1999)
[Publications] Bernier,N.J.,et al.: "Cardiovascular control via angiotensin II and circulating catecholamines in the spiny dogfish,Squalus acanthias." J.Comp.Physiol.(in press). (1999)
[Publications] Bernier,N.J.et al.: "Mediation of humoral catecholamine secretion by the renin-angiotensin system in hypotensive rainbow trout(Onchrhynchus mykiss)." J.Endocrinol.(in press). (1999)
[Publications] Tsuchida,Y.et al.: "Effects of homologous angiotensin II infusion at physiological doses on drinking and other thirst-related parameters in eels,Anguilla japonica." Zool.Sci.(in press). (1999)
[Publications] Cerra,M.C.et al.: "Localization and partial characterization of angiotensin II receptors in the heart of Scyliorhinus cannicula." Gen.Comp.Endocrinol.(in press). (1999)
[Publications] Hiruma,Y.et al.: "Endothelins inhibit mineralization of rat calvarial osteoblast-like cells." J.Cardiovasc.Pharmacol.31. S521-S523 (1998)
[Publications] Otsuka,E.et al.: "Effects of nitric oxide from exogenous nitric oxide donors on osteoblastic metabolism." Eur.J.Pharmacol.349. 345-350 (1999)

1998 Fiscal Year Annual Research Report

体液・循環調節に関与するペプチドホルモン受容体に関する比較分子生物学的研究(塩類細胞の分子解剖と分化誘導機構の解明)

Principal Investigator

広瀬 茂久 東京工業大学, 生命理工学部, 教授 (10134199)

Research Products

[Publications] Suzuki,Y.et al.: "Identification by differential display of a hypertonicity-inducible inward rectifier pottasium channel highly expressed in chloride cells." J.Biol.Chem.274(in press). (1999)

[Publications] Hagiwara,H.et al.: "Deceleration by angiotensin II of the differentiation and bone formation of rat calvarial osteoblastic cells." J.Endocrinol.156. 543-550 (1998)

[Publications] Hiruma,Y.et al.: "Endothelins inhibit the mineralization of osteoblastic MC3T3-E1 cells through thr A-type endothelin receptor." Am.J.Physiol.275. R1099-R1105 (1998)

[Publications] Kashiwagi,M.et al.: "Cloning,properties and tissue distribution of natriuretic peptide receptor-A of euryhaline eel,Anguilla japonica." Eur.J.Biochem.259. 204-211 (1999)

[Publications] Nagase,M.et al.: "Genomic organization and regulation of expression of the lectin-like oxidized low-density lipoprotein receptor(LOX-1)gene." J.Biol.Chem.273. 33702-33707 (1998)

[Publications] Tsuchida,T.et al.: "Effects of homologous atrial natriuretic peptide on drinking and plasma angiotensin II level in eels." Am.J.Physiol.230. R1605-R1610 (1998)

[Publications] Miyazaki,H.et al.: "Molecular cloning of CLC channels in Oreochromis mossambicus and their functional complementation of yeast CLC gene mutant" Biochem.Biophys.Res.Commun.(in press). (1999)

[Publications] Takei,Y.et al.: "Tetrapod-type[Asp1]angiotensin is present in a primitive holostean fish,Amia calva." Gen.Comp.Endocrinol.110. 140-146 (1998)

[Publications] Sakaguchi,H.et al.: "Characterisation of C-type natriuretic peptide receptors in the gill of dogfish Triakis scyllia." J.Endocrinol.156. 127-134 (1998)

[Publications] Tierney,M.L.et al.: "A radioimmunoassay for the detection of angiotensin II in elasmobranch fish." Gen.Comp.Endocrinol.111. 299-305 (1998)

[Publications] Takei,Y.et al.: "Antidiuretic effect of eel ANP infused at physiological doses in seawater-adapted eels,Anguilla japonica." Zool.Sci.15. 399-404 (1998)

[Publications] Suzuki,N.et al.: "Amino acid sequences of N-terminal procalcitonin of some vertebrates." Nether.J.Zool.48. 349-360 (1998)

[Publications] Hamano,K.et al.: "Direct vasoconstrictor action of homologous angiotensin II on isolated aterial ring preparation in an elasmobranch fish." J.Endocrinol.158. 419-423 (1997)

[Publications] McKendry,J.E.et al.: "The effects of netriuretic peptides on catecholamine release in American eel(Anguilla rostrata),rainbow trout(Oncorhynchus mykiss),and spiny dogfish(Squalus acanthias)." Fish Physiol.Biochem.(in press). (1999)

[Publications] Takei,Y.: "Comparative endocrinology of natriuretic peptide systems." Int.Rev.Cytol.(in press). (1999)

[Publications] Hazon,N.et al.: "he renin-angiotensin system in elasmobranch fish : A review." J.Exp.Zool.(in press). (1999)

[Publications] Takei,Y.: "Comparative physiology of body fluid regulation. in eels." Jpn.J.Physiol.(in press). (1999)

[Publications] Bernier,N.J.,et al.: "Cardiovascular control via angiotensin II and circulating catecholamines in the spiny dogfish,Squalus acanthias." J.Comp.Physiol.(in press). (1999)

[Publications] Bernier,N.J.et al.: "Mediation of humoral catecholamine secretion by the renin-angiotensin system in hypotensive rainbow trout(Onchrhynchus mykiss)." J.Endocrinol.(in press). (1999)

[Publications] Tsuchida,Y.et al.: "Effects of homologous angiotensin II infusion at physiological doses on drinking and other thirst-related parameters in eels,Anguilla japonica." Zool.Sci.(in press). (1999)

[Publications] Cerra,M.C.et al.: "Localization and partial characterization of angiotensin II receptors in the heart of Scyliorhinus cannicula." Gen.Comp.Endocrinol.(in press). (1999)

[Publications] Hiruma,Y.et al.: "Endothelins inhibit mineralization of rat calvarial osteoblast-like cells." J.Cardiovasc.Pharmacol.31. S521-S523 (1998)

[Publications] Otsuka,E.et al.: "Effects of nitric oxide from exogenous nitric oxide donors on osteoblastic metabolism." Eur.J.Pharmacol.349. 345-350 (1999)

広瀬茂久東京工業大学, 生命理工学部, 教授 (10134199)