1997 Fiscal Year Annual Research Report

神経情報伝達物質の放出制御の分子メカニズムの解明

Research Project

Project/Area Number	08044254
Research Institution	The University of Tokyo
Principal Investigator	御子柴克彦東京大学, 医科学研究所, 教授 (30051840)
Co-Investigator(Kenkyū-buntansha)	道川貴章東京大学, 医科学研究所, 助手 (90282516) 井上貴文東京大学, 医科学研究所, 助手 (10262081) 古市貞一東京大学, 医科学研究所, 助教授 (50219094) RODOLFO R.Li ニューヨーク大学, 医学部, 教授
Keywords	IP_4 / IP_5 / IP_6 / シナプトタグミン / 神経伝達物質放出 / イノシトールポリリン酸 / C2Aドメイン
Research Abstract	神経細胞にはこのIP3以外にも種々のイノシロールポリリン酸代謝産物が存在することからその重要性が考えられてきたが,特異的結合蛋白質が未道程であったため生理的機能の詳細は明らかではなかった。我々はイノシトールポリリン酸(IP_4,IP_5,IP_6)結合タンパク質として小脳よりシナプトタグミン精製することに成功した。シナプトタグミンの構造的特徴は膜貫通領域を一ケ所持ちC末端の細胞質側にはプロテインキナーゼCのC2調節領域とホモロジーを有する配列が2ケ所存在しそれぞれC2A,C2Bドメインと呼ばれている。C2Bドメインはカルシウム非依存的にリン脂質およびイノシトールポリリン酸(イノシトール-1、3、4、5、-四リン酸(IP_4)、イノシトール-1、3、4、5、6-五リン酸(IP_5)、イノシトール六リン酸(IP_6))を結合した。これらの結果から我々は二つのC2ドメインが神経伝達物質の放出の過程で異なる役割を持つものと考え、各々のドメインに対する特異的抵抗を作製した。次に我々はイカの巨大軸索のプレシナプスにこれらの抗体およびイノシトールポリリン酸(IP_4,IP_5,IP_6)をマイクロインジェクションすることにより以下の結論を得た。(i)シナプトタグミンのC2Aドメインは神経伝達物質放出の際のカルシウムセンサーとして機能しシナプス小胞とプレシナプス膜の融合の過程に関与すること、また(ii)C2Bドメインにはイノシトールポリリン酸が結合することによりC2Aドメインを介する膜融合の過程を阻害する(イノシトールポリリン酸が神経伝達物質放出の負の制御因子をして作用する)。さらに(iii)C2BドメインはClathrin assembly protein,AP-2を結合することによりシナプス小胞のリサイクリングの過程にも関与することが示唆された。

Research Products

(33 results)

All Other

All Publications (33 results)

[Publications] Del Rio, J. A: "A role for Cajal-Retzius cells and reelin in the development of hippocampal connections." Nature.385. 70-74 (1997)
[Publications] Umemori, H.: "Activation of the G Protein Gq/11 throuth tyrosine phosphorylation of the a subunit." Science.276. 1878-1882 (1997)
[Publications] Sheldon. M.: "Scrambmer and yotari disrupt the disabled gene and produce a reeler-like phenotype in mice" Nature.389. 730-733 (1997)
[Publications] Kume, S: "Role of Inositol 1, 4, 5-Trisphhosphate Receptor in Ventral Signaling in Xenopus Embryos." Science.278. 1940-1943 (1997)
[Publications] Zhao, H.: "Functional expression of a mammalian odorant receptor." Science.279. 237-242 (1998)
[Publications] D'Arcangelo, G: "Reelin is a seecred glycoprotein recognized by the CR-50 monoclonal antibody." J.Neurosci..17. 23-31 (1997)
[Publications] Ohara-ImaizumiM: "Distinct roles of C2A and C2B domains of synaptotagmin in the requlation of exocytosis in adrenal chromaffin cells." Proc. Natl. Acad. Sci. USA.94. 287-291 (1997)
[Publications] Kume, S: "Development Expression of the Inositol 1, 4, 5-Trisphosphate Receptor and Structural Changes in the Endoplasmic Reticulum during Oogenesis and Meiotic Maturation of Xenopus leavis." Developmental Biology.182.228-239 (1997)
[Publications] Mehrotra, B: "Selective photoaffinity lebeling of the inositiol polyphosphate binding C2B domains of synaptotagmins." J. Biol. Chem.272.4237-4244 (1997)
[Publications] Morita, N: "Astrocytic Lineage Analysis by Detection of GFAP Promoter Activity in vitro." Developmental Neuroscience.19.210-218 (1997)
[Publications] Sayars, L. G: "Intracellular targetting and homotetramer formation of a truncated inositol 1, 4, 5-trisphosphate receptor/green fluorescent protein chimera in Xenopus leavis oocytes." Biochem. J.323.273-280 (1997)
[Publications] Nagai, T.: "The expression of the mouse Zic1,Zic2,and Zic3 gene suggests an essential role forZic genes in body pattern formation." Developmental Biology.182.299-313 (1997)
[Publications] Yoneshima, H.: "Ca2_+ differetly regulates the ligand-affinity ststes of type 1 and type 3 inositol 1,4,5-trisphosphate receptors." Biochem. J.322.591-596 (1997)
[Publications] Tovey. S. C: "Identification and characerization of inositol 1, 4, 5-trisphosphate receptors in the rat testis." Cell Calcium.21.311-319 (1997)
[Publications] Hisatsume, C.: "Phosphorylation-dependent Regulation of N-Methy1-D-asparate Receptors by calmodulin." J. Biol. Chem.272.20805-20810 (1997)
[Publications] Maruyama, T.: "2APB, 2-aminoethoxydipphenyl borate, a membrance penetrable modulator of Ins (1, 4, 5) P3-induced Ca2_+ release." J. Biochem.122.498-505 (1997)
[Publications] Maruyama, T.: "Attenuation of intracellular Ca2_+ and secretory responses by Ins(1, 4, 5)P3-induced Ca2_+ release modulator, 2APB, In rat pancreatic acinar cells." Biomedical Research.18.297-302 (1997)
[Publications] Kume, S.: "Developmental expression of the inositol 1, 4, 5-trisphosphate receptor and localization of inositol 1, 4, 5-trisphosphate during early embryogenesis in Xenopus leavis." Mechanisms of Development.66.157-168 (1997)
[Publications] Tamura, H.: "High sulfotransferase activity for phenolic aromatic odorants present in the mouse olfactory." Chemico-Biological Interactions.104.1-9 (1997)
[Publications] Miyata, T.: "Regulation of Purkinje Cell Alignment by Reelin as Revealed with CR-50 Antibody." J. Neurosci.17.3599-3609 (1997)
[Publications] Nakayama, T: "Mapping of the Human HPC-1/Syntaxin 1A Gene (STX1A) to Chromosome 7 Band q11. 2." Genomics.42.173-176 (1997)
[Publications] Fukuda, M: "Regulation by bivalent cations of phospholipid bunding to the C2A domain of synaptotagmin III." Biochem. J.323. 421-425 (1997)
[Publications] Mochida, S.: "Roles of Synaptotagmin C2 Domains in Neurotransmitter secretion and inositol high-polyphospphate binding at mammalian cholinergic synapses." Neurosci.77. 937-943 (1997)
[Publications] Nakajima, K: "Distruption ofhippocampal development in vivo by CR-50 mAb against reelin." Proc. Natl. Acad. Sci.94. 8196-8201 (1997)
[Publications] Nakata, K.: "Xenopus Zic3, a primary regulator both in neural and neural crest development." Proc. Natl. Acad. Sci.94.11980-11985 (1997)
[Publications] Morikawa, K.: "Transcription initiation Sites and Promoter Structure of the Mouse Type 2 Inositol 1, 4, 5-Trisphospharate Receptor Gene." Gene.196.181-185 (1997)
[Publications] Wada, E.: "Generation and Characterization of Mice Lacking Gastrin-Releasing Peptide Receptor." Biochem. Biophys. Res. Comm.239.28-33 (1997)
[Publications] Taketo, M.: "Inositol-1, 3, 4, 5-tetrakisphosphate Binding Sites in Control and ras-Transformaed NIH/3T3 Fibroblasts." Biochem. Biophys. Res. Comm.239. 349-352 (1997)
[Publications] Lang. E. J.: "The first C2 domain of synaptotagmin is required for exocytosis of insulin from pancreatic b-cells : action of synaptotagmin at low micromolar calcium." EMBO J.16.5837-5846 (1997)
[Publications] Yoneshima, H.: "A novel neurological mutant mouse,yotari,which exhibits reeler-like phenotype but expresses CR-50 antigen/Reelin." Neuroscience Research.29.217-223 (1997)
[Publications] Konishi, Y: "Demonstration of an E-box and transcriptional regulation of the mouse type 1 Inositol 1, 4, 5-trisphosphate receptor gene." J. Neurochem.69. 476-484 (1997)
[Publications] Kojima, T.: "Characterization of the pleckstrin homology domain of Btk as an Inositol polyphosphate and phosphoinositide binding domain." Biochem. Biophys. Res. Comm.236. 333-339 (1997)
[Publications] Tamura, H: "High sulfotransferase activity for phenolic aromatic odorants present in the mouse olfactory organ." Chemico-Biological Interractions.104.1-9 (1997)

1997 Fiscal Year Annual Research Report

神経情報伝達物質の放出制御の分子メカニズムの解明

Principal Investigator

御子柴 克彦 東京大学, 医科学研究所, 教授 (30051840)

Research Products

[Publications] Del Rio, J. A: "A role for Cajal-Retzius cells and reelin in the development of hippocampal connections." Nature.385. 70-74 (1997)

[Publications] Umemori, H.: "Activation of the G Protein Gq/11 throuth tyrosine phosphorylation of the a subunit." Science.276. 1878-1882 (1997)

[Publications] Sheldon. M.: "Scrambmer and yotari disrupt the disabled gene and produce a reeler-like phenotype in mice" Nature.389. 730-733 (1997)

[Publications] Kume, S: "Role of Inositol 1, 4, 5-Trisphhosphate Receptor in Ventral Signaling in Xenopus Embryos." Science.278. 1940-1943 (1997)

[Publications] Zhao, H.: "Functional expression of a mammalian odorant receptor." Science.279. 237-242 (1998)

[Publications] D'Arcangelo, G: "Reelin is a seecred glycoprotein recognized by the CR-50 monoclonal antibody." J.Neurosci..17. 23-31 (1997)

[Publications] Ohara-ImaizumiM: "Distinct roles of C2A and C2B domains of synaptotagmin in the requlation of exocytosis in adrenal chromaffin cells." Proc. Natl. Acad. Sci. USA.94. 287-291 (1997)

[Publications] Kume, S: "Development Expression of the Inositol 1, 4, 5-Trisphosphate Receptor and Structural Changes in the Endoplasmic Reticulum during Oogenesis and Meiotic Maturation of Xenopus leavis." Developmental Biology.182.228-239 (1997)

[Publications] Mehrotra, B: "Selective photoaffinity lebeling of the inositiol polyphosphate binding C2B domains of synaptotagmins." J. Biol. Chem.272.4237-4244 (1997)

[Publications] Morita, N: "Astrocytic Lineage Analysis by Detection of GFAP Promoter Activity in vitro." Developmental Neuroscience.19.210-218 (1997)

[Publications] Sayars, L. G: "Intracellular targetting and homotetramer formation of a truncated inositol 1, 4, 5-trisphosphate receptor/green fluorescent protein chimera in Xenopus leavis oocytes." Biochem. J.323.273-280 (1997)

[Publications] Nagai, T.: "The expression of the mouse Zic1,Zic2,and Zic3 gene suggests an essential role forZic genes in body pattern formation." Developmental Biology.182.299-313 (1997)

[Publications] Yoneshima, H.: "Ca2_+ differetly regulates the ligand-affinity ststes of type 1 and type 3 inositol 1,4,5-trisphosphate receptors." Biochem. J.322.591-596 (1997)

[Publications] Tovey. S. C: "Identification and characerization of inositol 1, 4, 5-trisphosphate receptors in the rat testis." Cell Calcium.21.311-319 (1997)

[Publications] Hisatsume, C.: "Phosphorylation-dependent Regulation of N-Methy1-D-asparate Receptors by calmodulin." J. Biol. Chem.272.20805-20810 (1997)

[Publications] Maruyama, T.: "2APB, 2-aminoethoxydipphenyl borate, a membrance penetrable modulator of Ins (1, 4, 5) P3-induced Ca2_+ release." J. Biochem.122.498-505 (1997)

[Publications] Maruyama, T.: "Attenuation of intracellular Ca2_+ and secretory responses by Ins(1, 4, 5)P3-induced Ca2_+ release modulator, 2APB, In rat pancreatic acinar cells." Biomedical Research.18.297-302 (1997)

[Publications] Kume, S.: "Developmental expression of the inositol 1, 4, 5-trisphosphate receptor and localization of inositol 1, 4, 5-trisphosphate during early embryogenesis in Xenopus leavis." Mechanisms of Development.66.157-168 (1997)

[Publications] Tamura, H.: "High sulfotransferase activity for phenolic aromatic odorants present in the mouse olfactory." Chemico-Biological Interactions.104.1-9 (1997)

[Publications] Miyata, T.: "Regulation of Purkinje Cell Alignment by Reelin as Revealed with CR-50 Antibody." J. Neurosci.17.3599-3609 (1997)

[Publications] Nakayama, T: "Mapping of the Human HPC-1/Syntaxin 1A Gene (STX1A) to Chromosome 7 Band q11. 2." Genomics.42.173-176 (1997)

[Publications] Fukuda, M: "Regulation by bivalent cations of phospholipid bunding to the C2A domain of synaptotagmin III." Biochem. J.323. 421-425 (1997)

[Publications] Mochida, S.: "Roles of Synaptotagmin C2 Domains in Neurotransmitter secretion and inositol high-polyphospphate binding at mammalian cholinergic synapses." Neurosci.77. 937-943 (1997)

[Publications] Nakajima, K: "Distruption ofhippocampal development in vivo by CR-50 mAb against reelin." Proc. Natl. Acad. Sci.94. 8196-8201 (1997)

[Publications] Nakata, K.: "Xenopus Zic3, a primary regulator both in neural and neural crest development." Proc. Natl. Acad. Sci.94.11980-11985 (1997)

[Publications] Morikawa, K.: "Transcription initiation Sites and Promoter Structure of the Mouse Type 2 Inositol 1, 4, 5-Trisphospharate Receptor Gene." Gene.196.181-185 (1997)

[Publications] Wada, E.: "Generation and Characterization of Mice Lacking Gastrin-Releasing Peptide Receptor." Biochem. Biophys. Res. Comm.239.28-33 (1997)

[Publications] Taketo, M.: "Inositol-1, 3, 4, 5-tetrakisphosphate Binding Sites in Control and ras-Transformaed NIH/3T3 Fibroblasts." Biochem. Biophys. Res. Comm.239. 349-352 (1997)

[Publications] Lang. E. J.: "The first C2 domain of synaptotagmin is required for exocytosis of insulin from pancreatic b-cells : action of synaptotagmin at low micromolar calcium." EMBO J.16.5837-5846 (1997)

[Publications] Yoneshima, H.: "A novel neurological mutant mouse,yotari,which exhibits reeler-like phenotype but expresses CR-50 antigen/Reelin." Neuroscience Research.29.217-223 (1997)

[Publications] Konishi, Y: "Demonstration of an E-box and transcriptional regulation of the mouse type 1 Inositol 1, 4, 5-trisphosphate receptor gene." J. Neurochem.69. 476-484 (1997)

[Publications] Kojima, T.: "Characterization of the pleckstrin homology domain of Btk as an Inositol polyphosphate and phosphoinositide binding domain." Biochem. Biophys. Res. Comm.236. 333-339 (1997)

[Publications] Tamura, H: "High sulfotransferase activity for phenolic aromatic odorants present in the mouse olfactory organ." Chemico-Biological Interractions.104.1-9 (1997)

御子柴克彦東京大学, 医科学研究所, 教授 (30051840)