1999 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, 助教授 (10129249)
Keywords	循環調節 / 内向き整流性Kチャネル / ナトリウム利尿ペプチド / 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の解明に新視点を導入するべく,比較分子生物学的なアプローチを試みつつある。本年度は,淡水・海水の両方に適応できるウナギと高度な酸性適応能を有する恐山ウグイの系を中心に以下の成果を得た。1)ウナギで発見した新規Kチャネルの哺乳類型(Kir7.1)の生理機能:Kir7.1の組織分布及び細胞内分布の極性(apical vs basolateral)がNa,K-ATPaseのものとよく一致することから,Kir7.1が生理学上極めて重要で,古くから探し求められていたNa,K-ATPaseのパートナー分子である可能性が強いことを示唆した。2)pH3.5という強酸性下で生きている恐山ウグイの耐酸性機構を分子レベルである程度説明できるようにした。すなわちNa/Hexchangerをクローニングし,その発現が酸性下で顕著に高まり,かつ塩類細胞のapical膜に局在することを抗体染色によって示した。3)塩類細胞の分化誘導に関わると期待されるRBCCタンパク質を同定することに成功した。4)密度勾配遠心法を用いて塩類細胞を50%以上に濃縮する技術を確立した。その結果,細胞レベルでイオンや細胞内情報伝達系の変化をリアルタイムで調べられるようになった。5)^<22>Naを用いてNaイオンの出入りを調べ,淡水ウナギではCNPがNaの取り込みを上昇させ,海水中では逆に排出を促進すること,また単離したエラ上皮細胞にCNPを投与するとセカンドメッセンジャーであるcGMPが増加することを示した。7)ナトリウム利尿ペプチド受容体に対する抗体を種々作成し,免疫組織化学的解析を行い,CNPに特異的なB型受容体が血管平滑筋に多く発現していることを明らかにした。CNPはエラにおける血流量を調節することによりイオンの出入りを調節している可能性が強い。

Research Products

(20 results)

All Other

All Publications (20 results)

[Publications] Abe, J. et al.: "Ig-Hepta, a novel member of the G protein-coupled hepta-helical receptor (GPCR) family that has immunoglobulin-like repeats in a long N-terminal extracellular domain and defines a new subfamily of GPCRs"J. Biol. Chem.. 247. 19957-19964 (1999)
[Publications] Hagiwara, H. et al.: "Ipriflavone down-regulates endothelin receptor levels during differentiation of rat calvarial osteoblast-like cells"J. Biochem.. 126. 168-173 (1999)
[Publications] Honda, S. et al.: "Regulation by la,25-dihydroxyviatmin D3 of expression of stanniocalcin message in the rat kidney and ovary"FEBS Lett.. 459. 119-122 (1999)
[Publications] Inoue, A. et al.: "Correlation between induction of expression of biglycan and mineralization by C-type matriuretic peptide in osteoblastic cells"J. Biochem.. 125. 103-108 (1999)
[Publications] Massareli, E.E. et al.: "Differential subcellular distribution of neurolysin (EC 3.4.24.16) and thimet oligopeptidase (EC 3.4.24.15) in the rat brain"Brain Res.. 851. 261-265 (1999)
[Publications] Nagase, M. et al.: "Retrotransposons transcribed preferentially in proximal tubules of salt-hypertensive rat"Kidney International. 55. 995-1004 (199)
[Publications] Nakamura, N. et al.: "Inwardly rectifying potassium channel kir7.1 is highly expressed in the thyroid follicular cells, intestinal epithelial cells and choroid plexus epithelial cells : implication for a functional coupling with Na^+, K^+-ATPase"Biochem. J.. 342. 329-336 (1999)
[Publications] Otsuka, E. et al.: "Characterization of osteoblastic differentiation of stromalcell line ST2 that is induced by ascorbic acid"Am. J. Physiol.. 277. C132-C138 (1999)
[Publications] Schalkwijk, J. et al.: "The trappin gene family : proteins defined by an N-terminal transglutaminase substrate domain and a C-terminal four-disulfide core"Biochem. J.. 259. 569-577 (1999)
[Publications] 竹井祥郎: "比較することの意味と面白さ(講談社ブルーバックス)"ホルモンから見た生命 (日本比較内分泌学会編). (1999)
[Publications] McKendry, J.E. et al.: "Natriuretic peptides and the control of catecholamine release in two freshwater teleost and a marine elasmobranch fish"Fish Physiol. Biochem.. 20. 61-77 (1999)
[Publications] Joss, J.M.P. et al.: "Teleost-type angiotensin is present in Australian lungfish, Neoceratodus forsteri"Gen. Comp. Endocrinol.. 114. 206-212 (1999)
[Publications] Tsuchida, T., & Takei, Y.: "A potent dipsogenic action of homologous angiotensin II infused at physiological doses in eels, Anguilla japonica"Zool. Sci.. 16. 479-483 (1999)
[Publications] Cerra, M.C. et al.: "Localization and partial characterization and partial characterization of angiotensin II receptors in the heart of Scyliorhinus cannicula. brain of a teleost, stargazer, Uranoscops japonicus"Gen. Comp. Endocrinol.. (in press). (2000)
[Publications] Takei, Y. et al.: "Interaction of the natriuretic peptide system and the renin-angiotensin system in eel osmoregulation"Recent Development in Comparative Endocrinology and Neurobiology (E.W. Roubes et al. eds.). 144-148 (1999)
[Publications] Tsuchida, T., & Takei, Y.: "Interaction of ANP and ANGII in regulation of drinking in eels"Recent Progress n Molecular and Comparative Endocrinology (H.B. Kwon et al. eds.). 453-459 (1999)
[Publications] Tsuchida, T., & Takei, Y.: "A potent disogenic action of homologous antiotensin II infused at physiological doses in eels. Anguilla japonica"Zool. Sci.. 16. 479-483 (1999)
[Publications] Takei, Y.: "Structural and functional evolution of the matriuretic peptide system in vertebrates"Int. Rev. Cytol.. 194. 1-66 (2000)
[Publications] Takei, Y.: "Comparative physiology of body fluid regulation in vertebrates with special reference to thirst regulation"Jpn. J. Physiol.. (in press). (2000)
[Publications] Tsuchida, T., & Takei, Y.: "Peripheral renin-angiotensin system in not involved in vigorous drinking of seawater-adapted eels, Anguilla japonica"Am. J. Physiol.. (in press). (2000)

1999 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] Abe, J. et al.: "Ig-Hepta, a novel member of the G protein-coupled hepta-helical receptor (GPCR) family that has immunoglobulin-like repeats in a long N-terminal extracellular domain and defines a new subfamily of GPCRs"J. Biol. Chem.. 247. 19957-19964 (1999)

[Publications] Hagiwara, H. et al.: "Ipriflavone down-regulates endothelin receptor levels during differentiation of rat calvarial osteoblast-like cells"J. Biochem.. 126. 168-173 (1999)

[Publications] Honda, S. et al.: "Regulation by la,25-dihydroxyviatmin D3 of expression of stanniocalcin message in the rat kidney and ovary"FEBS Lett.. 459. 119-122 (1999)

[Publications] Inoue, A. et al.: "Correlation between induction of expression of biglycan and mineralization by C-type matriuretic peptide in osteoblastic cells"J. Biochem.. 125. 103-108 (1999)

[Publications] Massareli, E.E. et al.: "Differential subcellular distribution of neurolysin (EC 3.4.24.16) and thimet oligopeptidase (EC 3.4.24.15) in the rat brain"Brain Res.. 851. 261-265 (1999)

[Publications] Nagase, M. et al.: "Retrotransposons transcribed preferentially in proximal tubules of salt-hypertensive rat"Kidney International. 55. 995-1004 (199)

[Publications] Nakamura, N. et al.: "Inwardly rectifying potassium channel kir7.1 is highly expressed in the thyroid follicular cells, intestinal epithelial cells and choroid plexus epithelial cells : implication for a functional coupling with Na^+, K^+-ATPase"Biochem. J.. 342. 329-336 (1999)

[Publications] Otsuka, E. et al.: "Characterization of osteoblastic differentiation of stromalcell line ST2 that is induced by ascorbic acid"Am. J. Physiol.. 277. C132-C138 (1999)

[Publications] Schalkwijk, J. et al.: "The trappin gene family : proteins defined by an N-terminal transglutaminase substrate domain and a C-terminal four-disulfide core"Biochem. J.. 259. 569-577 (1999)

[Publications] 竹井祥郎: "比較することの意味と面白さ(講談社ブルーバックス)"ホルモンから見た生命 (日本比較内分泌学会編). (1999)

[Publications] McKendry, J.E. et al.: "Natriuretic peptides and the control of catecholamine release in two freshwater teleost and a marine elasmobranch fish"Fish Physiol. Biochem.. 20. 61-77 (1999)

[Publications] Joss, J.M.P. et al.: "Teleost-type angiotensin is present in Australian lungfish, Neoceratodus forsteri"Gen. Comp. Endocrinol.. 114. 206-212 (1999)

[Publications] Tsuchida, T., & Takei, Y.: "A potent dipsogenic action of homologous angiotensin II infused at physiological doses in eels, Anguilla japonica"Zool. Sci.. 16. 479-483 (1999)

[Publications] Cerra, M.C. et al.: "Localization and partial characterization and partial characterization of angiotensin II receptors in the heart of Scyliorhinus cannicula. brain of a teleost, stargazer, Uranoscops japonicus"Gen. Comp. Endocrinol.. (in press). (2000)

[Publications] Takei, Y. et al.: "Interaction of the natriuretic peptide system and the renin-angiotensin system in eel osmoregulation"Recent Development in Comparative Endocrinology and Neurobiology (E.W. Roubes et al. eds.). 144-148 (1999)

[Publications] Tsuchida, T., & Takei, Y.: "Interaction of ANP and ANGII in regulation of drinking in eels"Recent Progress n Molecular and Comparative Endocrinology (H.B. Kwon et al. eds.). 453-459 (1999)

[Publications] Tsuchida, T., & Takei, Y.: "A potent disogenic action of homologous antiotensin II infused at physiological doses in eels. Anguilla japonica"Zool. Sci.. 16. 479-483 (1999)

[Publications] Takei, Y.: "Structural and functional evolution of the matriuretic peptide system in vertebrates"Int. Rev. Cytol.. 194. 1-66 (2000)

[Publications] Takei, Y.: "Comparative physiology of body fluid regulation in vertebrates with special reference to thirst regulation"Jpn. J. Physiol.. (in press). (2000)

[Publications] Tsuchida, T., & Takei, Y.: "Peripheral renin-angiotensin system in not involved in vigorous drinking of seawater-adapted eels, Anguilla japonica"Am. J. Physiol.. (in press). (2000)

広瀬茂久東京工業大学, 生命理工学部, 教授 (10134199)