1996 Fiscal Year Annual Research Report

消化管粘膜上皮細胞における細胞内情報伝達機構の解析

Research Project

Project/Area Number	08044331
Research Institution	University of Shizuoka
Principal Investigator	桑原厚和静岡県立大学, 大学院, 助教授 (60142890)
Co-Investigator(Kenkyū-buntansha)	RECHKEMMER ジードイツ国立栄養生理学研究所, 教授 LABURTHE M. フランス国立衛生学研究所, 部長杉谷博士日本大学松戸歯学部, 助教授 (20050114) 望月徹静岡県立大学, 薬学部, 助教授 (00117780) 藤宮峯子滋賀医科大学, 医学部, 助教授 (10199359)
Keywords	Guinea-pig / Distal Colon / Chloride Secretion / Cytosolic Calcium / Tachykinin / Crypt Cell / Conforcal microscopy / Ion Transport
Research Abstract	以前採択された国際学術共同研究により神経ペプチドが消化管粘膜上皮細胞からの水分泌を誘発し、その作用はそれぞれのペプチドの前処置により著明に増強されることを観察した。これらペプチドは細胞内情報伝達系としてカルシウム及び環状ヌクレオチドを利用すると考えられるが、現在その作用機構については明らかではない。本年度は、細胞内情報伝達系としてカルシウムイオンを利用すると考えられるSubstancePによるクロライドイオン分泌の細胞内情報伝達機構をモルモット下部大腸より単離したCrypt cellを使用して、共焦点レーザー顕微鏡により解析した。 substance P、Neurokinin AおよびNeurokinin Bの適用により、Crypt cellsの[Ca^<2+>]_iは用量依存症に増加し、その[Ca^<2+>]_iの上昇は一過性であった。またその反応性は、SubstanceP>NeurokininA>>NeurokininBの順であった。選択的NK-1受容体拮抗薬、FK888の前処置により、SubstancePおよびNeurokinkin Aによる[Ca^<2+>]_iの上昇は有意に抑制された。また、細胞外のCa^<2+>を除去した場合、SubstancePによる[Ca^<2+>]_i上昇の頂値は、変化しなかったが、変化の積分により算出した総[Ca^<2+>]_i変化量が減少した。またThapsigarginの前処理によっても、SubstancePによる[Ca^<2+>]_iの頂値および総[Ca^<2+>]_i変化量は有意に抑制されたが、完全には抑制しなかった。Thapsigargin処理と細胞外からCa^<2+>を除去処理した場合、SubstancePによる[Ca^<2+>]_iの頂値および総[Ca^<2+>]_i変化量が著しく抑制された。これらの実験結果から、SubstancePによる[Ca^<2+>]_iの上昇は、初期に細胞内Ca^<2+>貯蔵部位からCa^<2+>が遊離し、続いて細胞外からCa^<2+>が流入することが考えられた。さらにUssing Chamberを用いた実験で、基底膜側(漿膜側)のCa^<2+>除去により、SubstancePによるクロライドイオン分泌は完全に抑制された。しかしながら、頂端膜側(粘膜側)Ca^<2+>除去は、SubstancePによるクロライドイオン分泌に影響を与えなかった。従って、細胞外からのCa^<2+>流入は、主として基底膜側から流入していると考えられた。以上の結果から、粘膜上皮Crypt cellsにおけるSubstancePによるクロライドイオン分泌機序には、小胞体からのCa^<2+>の放出と同時に、細胞外からのCa^<2+>流入の関与が示唆され、特に、基底膜側からのCa^<2+>の流入が重要な役割を有していることが示唆された。

Research Products
(21 results)

All Other

All Publications (21 results)

[Publications] Fujimiya, M., Okumiya, K. and Kuwahara, A.: "Immunoelectron microscopic demonstration of Iuminal release of serotonin from rat enterochromaffin cells induced by high intraluminal pressure" Histochemistry and Cell Biology. (in press). (1997)
[Publications] Kakuyama, H., Kuwahara, A., Mochizuki, T., Hoshino, M. and Yanaihara, Y.: "Role of N-terminal active sites of galanin on neurally-evoked ciruclar muscle contractions in the guinea-pig ileum" Eur. J. Pharmacol.(in press). (1997)
[Publications] Kadowaki, M., Gershon, M.D. and Kuwahara, A: "Is nitric oxide involved in 5-HT-induced fluid secre-tion in the gut7" Behavioural Brain Research. 73(1/2). 293-296 (1996)
[Publications] Kuwahara, A. and Kadowaki M.: "Adenosine modulates PACAP-evoked chloride secretion in guinea-pig distal colon" Regulatory Peptides. 64. 101 (1996)
[Publications] Nagahama, M., Tsuzuki, T., Kuwahara, A. and MOchizuki, T.: "Light and electron microscopical studies of PACAP(pituitary adenylate cyclase activating polypeptide)immunoreactive neurons in the myenteric plexus of rat intestine" Regulatory Peptides. 64. 137 (1996)
[Publications] Kakuyama, H., Kuwahara, A., Mochizuki, T., Hoshino, M. and Yanaihara, N.: "Inhibitory effect of galanin(1-15)ーol analogs on neurally-evoked circular muscle contraction in the guinea-pig ileum" Regulatory Peptides. 64. 84 (1996)
[Publications] Fujimaiya, M. and Kuwahara, A: "Effect of CCK, VIP, PACAP27, 38 on release of serotonin from perfused rat intestine" Regulatory Peptides. 64. 48 (1996)
[Publications] Fujimiya, M., Okumiya, K. and Kuwahara, A.: "Immuno-electronmicroscopic demonstration of Iuminal release of serotonin stimulated by high pressure" Regulatory Peptides. 64. 146 (1996)
[Publications] Yamamoto, I., Kuwahara, A., Fujimura, M., Kadowaki, M., and Fujimiya, M.: "Involvement of 5-HT_3,5-HT_4 receptors on the motor activity of rat duodenum" Regulatory Peptides. 64. 212 (1996)
[Publications] Niisato, N., Ogata, Y., Nakao, S., Furuyama, S. and Sugiya, H.: "Bradykinin regulates histamine-induced Ca^<2+> mobilization via protein kinase C activation in human gingival fibroblasts" Cell Calcium. (in press). (1997)
[Publications] Ogata, Y., Niisato, N.,Furuyama, S., Cheifetz, S., Kim, R.H., Sugiya, H. and Sodek, J.: "Transforming growth factor-β1 regulation of born sialoprotein (BSP)gene transcription. Identification of a TGF-β activation element(TAE)in the rat BSP gene promoter" J.Cell Biochem.(in press). (1997)
[Publications] Ogata, Y., Niisato, N., Moriwaki, K., Yokota,Y. Furuyama, S. and Sugiya, H.: "Cementum, root dentin and born extracts stimulate chemotactic behavior in cells from periodontal tissue" Copm. Biochem. Physiol.(in press). (1997)
[Publications] Niisato, N., Ogata, Y., Furuyama, S. and Sugiya, H.: "Histamine H_1 receptor-induced Ca^<2+> mobilization and prostaglandin E_2 release in human gingival fibroblasts" Biochem. Pharmachol.,. 52. 1015-1023 (1996)
[Publications] Yokoyama, N., Kobayashi, T., Tamura, S. and Sugiya, H.: "Purification and characterization of protein phosphastase C in rat parotid acinar cells: two forms Mg^<2+>-activated histon phosphatase and phosphorylation by cAMPーdependent protein kinase" Arch. Biochem. Biophys.331(1). 1-8 (1996)
[Publications] Fujita-Yoshigaki, J., Dohke, Y., Hara-Yokoyama, M., Kamata, Y., Kozaki, S. Furuyama, S. and Sugiya, H.: "Vesicle-associated membrane protein 2 is essential for cAMP-regulated exocytosis in rat parotid acinar cells" J. Biol. Chem.271(22). 13130-13134 (1996)
[Publications] Hara-Yokoyama, M., Kukimoto I., Nishina, H., Kontani, K., Hirabayashi, Y., Irei, F., Sugiya, H., Furuyama, S. and Katada, T.: "Inhibition of NAD^+ glycohydrolase and ADP-ribosyl cyclase activities of leukocyte cell surface antigen CD38 by gangliosides" J. Biol. Chem.271(22). 12951-12955 (1996)
[Publications] Niisato, N., Ogata, Y., Furuyama, S. and Sugiya, H.: "Histamine H_1 receptor-stimulated Ca^<2+> signaling pathway in human periodontal ligament cells" J. Periodontal. Res.31. 113-119 (1996)
[Publications] Okumiya, K., Matssubayashi, K., Maeda, T. and Fujimiya, M.: "Changes in subcellular localization of Gastrin like immunoreactivity in epithelial cells of rat duodenum induced by carbachol" Peptides. 17. 225-232 (1996)
[Publications] Tadokoro, C., Yoshimoto, Y., Sakata, M., Fujimiya, M., Kuruchi, H., Adachi, E., Maeda, T. and Miyake, A.: "Localization of human placental glucose transporter 1 during pregnancy. An immunohistochemical study" Histology and Histopathology. 11. 673-681 (1996)
[Publications] Fujimiya, M., Okumiya, K., Yamane, T. and Maeda, T.: "Distribution of serotonin-immunoreactive nerve cells and fibers in the rat gadstrointestinal tract" Histochemistry and Cell Biology. 107. 105-114 (1997)
[Publications] 藤宮峯子・山本育男桑原厚和: "ラット十二指腸環流法による血液及び管腔中へのセロトニン分泌の研究-特に腸管内圧亢進の両分泌に与える影響について-" 消化管ホルモン. XIV. 70-75 (1996)

1996 Fiscal Year Annual Research Report

消化管粘膜上皮細胞における細胞内情報伝達機構の解析

Principal Investigator

桑原 厚和 静岡県立大学, 大学院, 助教授 (60142890)

Research Products

[Publications] Fujimiya, M., Okumiya, K. and Kuwahara, A.: "Immunoelectron microscopic demonstration of Iuminal release of serotonin from rat enterochromaffin cells induced by high intraluminal pressure" Histochemistry and Cell Biology. (in press). (1997)

[Publications] Kakuyama, H., Kuwahara, A., Mochizuki, T., Hoshino, M. and Yanaihara, Y.: "Role of N-terminal active sites of galanin on neurally-evoked ciruclar muscle contractions in the guinea-pig ileum" Eur. J. Pharmacol.(in press). (1997)

[Publications] Kadowaki, M., Gershon, M.D. and Kuwahara, A: "Is nitric oxide involved in 5-HT-induced fluid secre-tion in the gut7" Behavioural Brain Research. 73(1/2). 293-296 (1996)

[Publications] Kuwahara, A. and Kadowaki M.: "Adenosine modulates PACAP-evoked chloride secretion in guinea-pig distal colon" Regulatory Peptides. 64. 101 (1996)

[Publications] Nagahama, M., Tsuzuki, T., Kuwahara, A. and MOchizuki, T.: "Light and electron microscopical studies of PACAP(pituitary adenylate cyclase activating polypeptide)immunoreactive neurons in the myenteric plexus of rat intestine" Regulatory Peptides. 64. 137 (1996)

[Publications] Kakuyama, H., Kuwahara, A., Mochizuki, T., Hoshino, M. and Yanaihara, N.: "Inhibitory effect of galanin(1-15)ーol analogs on neurally-evoked circular muscle contraction in the guinea-pig ileum" Regulatory Peptides. 64. 84 (1996)

[Publications] Fujimaiya, M. and Kuwahara, A: "Effect of CCK, VIP, PACAP27, 38 on release of serotonin from perfused rat intestine" Regulatory Peptides. 64. 48 (1996)

[Publications] Fujimiya, M., Okumiya, K. and Kuwahara, A.: "Immuno-electronmicroscopic demonstration of Iuminal release of serotonin stimulated by high pressure" Regulatory Peptides. 64. 146 (1996)

[Publications] Yamamoto, I., Kuwahara, A., Fujimura, M., Kadowaki, M., and Fujimiya, M.: "Involvement of 5-HT_3,5-HT_4 receptors on the motor activity of rat duodenum" Regulatory Peptides. 64. 212 (1996)

[Publications] Niisato, N., Ogata, Y., Nakao, S., Furuyama, S. and Sugiya, H.: "Bradykinin regulates histamine-induced Ca^<2+> mobilization via protein kinase C activation in human gingival fibroblasts" Cell Calcium. (in press). (1997)

[Publications] Ogata, Y., Niisato, N., Moriwaki, K., Yokota,Y. Furuyama, S. and Sugiya, H.: "Cementum, root dentin and born extracts stimulate chemotactic behavior in cells from periodontal tissue" Copm. Biochem. Physiol.(in press). (1997)

[Publications] Niisato, N., Ogata, Y., Furuyama, S. and Sugiya, H.: "Histamine H_1 receptor-induced Ca^<2+> mobilization and prostaglandin E_2 release in human gingival fibroblasts" Biochem. Pharmachol.,. 52. 1015-1023 (1996)

[Publications] Fujita-Yoshigaki, J., Dohke, Y., Hara-Yokoyama, M., Kamata, Y., Kozaki, S. Furuyama, S. and Sugiya, H.: "Vesicle-associated membrane protein 2 is essential for cAMP-regulated exocytosis in rat parotid acinar cells" J. Biol. Chem.271(22). 13130-13134 (1996)

[Publications] Niisato, N., Ogata, Y., Furuyama, S. and Sugiya, H.: "Histamine H_1 receptor-stimulated Ca^<2+> signaling pathway in human periodontal ligament cells" J. Periodontal. Res.31. 113-119 (1996)

[Publications] Okumiya, K., Matssubayashi, K., Maeda, T. and Fujimiya, M.: "Changes in subcellular localization of Gastrin like immunoreactivity in epithelial cells of rat duodenum induced by carbachol" Peptides. 17. 225-232 (1996)

[Publications] Tadokoro, C., Yoshimoto, Y., Sakata, M., Fujimiya, M., Kuruchi, H., Adachi, E., Maeda, T. and Miyake, A.: "Localization of human placental glucose transporter 1 during pregnancy. An immunohistochemical study" Histology and Histopathology. 11. 673-681 (1996)

[Publications] Fujimiya, M., Okumiya, K., Yamane, T. and Maeda, T.: "Distribution of serotonin-immunoreactive nerve cells and fibers in the rat gadstrointestinal tract" Histochemistry and Cell Biology. 107. 105-114 (1997)

[Publications] 藤宮峯子・山本育男 桑原厚和: "ラット十二指腸環流法による血液及び管腔中へのセロトニン分泌の研究-特に腸管内圧亢進の両分泌に与える影響について-" 消化管ホルモン. XIV. 70-75 (1996)

桑原厚和静岡県立大学, 大学院, 助教授 (60142890)

[Publications] 藤宮峯子・山本育男桑原厚和: "ラット十二指腸環流法による血液及び管腔中へのセロトニン分泌の研究-特に腸管内圧亢進の両分泌に与える影響について-" 消化管ホルモン. XIV. 70-75 (1996)