情報伝達におけるGTP結合蛋白の役割に関する卵母細胞を用いた分子神経生物学的研究

利用卵母细胞研究 GTP 结合蛋白在信息传递中的作用的分子神经生物学研究

• 批准号: 03454492
• 负责人: SATOH Masamichi
• 金额: $ 4.54万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03454492/
• 关键词: Xenopus oocyte; mRNA; antisense DNA; G-protein; GTP-binding; muscarinic receptor; serotonin receptor; hybrid arrest; ムスカリンレセプタ-; セロトニンレセプタ-; ホスホリパ-ゼC; ラット脳

To Assign GTP-binding protein (G-protein) subtyps involved in the signal transduction from exogenous receptors to phospholipase C in the Xenopus oocyte translation system, antisense DNA complementary to rat G-protein alpha-subunit mRNA was designed and injected together with rat brain poly(A)^+mRNA. Current responses of mRNA-injected oocytes to acetylcholine (ACh) was suppressed by a co-injection of Gilalpha-antisense DNA dose-dependently, but response of the same oocytes to serotonin (5-HT) was not inhibited. In the oocytes co-injected with Goalpha-antisense DNA, the 5-HT response was more effectively suppressed than the ACh response. These result suggest that Goalpha but not Gilalpha intermediates brain 5-HTlc receptor function, and in contrast, muscarinic receptors derived from rat brain utilize Gilalpha rather than Goalpha to activate phospholipase C. Putative effects of G-protein on voltage-dependent calcium channels were also suggested.

为了确定非洲爪蟾卵母细胞翻译系统中参与外源性受体向磷脂酶C信号转导的GTP结合蛋白（G蛋白）亚型，设计了与大鼠G蛋白α亚基mRNA互补的反义DNA，并将其与大鼠脑poly（A）^+mRNA一起注射。注射mRNA的卵母细胞对乙酰胆碱（ACh）的电流反应被共注射Gilalpha-反义DNA剂量依赖性地抑制，但相同的卵母细胞对5-羟色胺（5-HT）的反应不受抑制。在卵母细胞共注射Goalpha-反义DNA，5-HT反应比ACh反应更有效地抑制。这些结果表明，Goalpha而不是Gilalpha介导脑5-HTlc受体功能，相反，来自大鼠脑的毒蕈碱受体利用Gilalpha而不是Goalpha来激活磷脂酶C。还提出了G蛋白对电压依赖性钙通道的推定作用。

项目成果

金子 周司(遠山 正彌監修): "アフリカツメガエル卵母細胞発現系ー電位固定法による膜蛋白発現の検出 「神経科学研究の先端技術プロトコルII)" 厚生社, 11 (1992)

Shuji Kaneko（由 Masaya Toyama 监督）：“非洲爪蟾卵母细胞表达系统 - 通过电压钳法检测膜蛋白表达‘神经科学研究先进技术方案 II)’” Koseisha, 11 (1992)

Shuji Kaneko, Hideki Takahashi and Masamichi Satoh: "Metabotropic responses to acetylcholine and serotonin of Xenopus oocytes injected with rat brain mRNA are transduced by different G-protein subtypes" FEBS Lett.299. 179-182 (1992)

Shuji Kaneko、Hideki Takahashi 和 Masamichi Satoh：“注射了大鼠脑 mRNA 的非洲爪蟾卵母细胞对乙酰胆碱和血清素的代谢反应是由不同的 G 蛋白亚型转导的”FEBS Lett.299。

Shuji Kaneko: "Metabotropic responses to acetylcholine and serotonin of Xenopus oocytes injected with rat brain mRNA are transduced by different G-protein subtypes" FEBS Lett.299. 179-182 (1992)

Shuji Kaneko：“注射了大鼠脑 mRNA 的爪蟾卵母细胞对乙酰胆碱和血清素的代谢反应是由不同的 G 蛋白亚型转导的”FEBS Lett.299。

Shuji Kaneko: "Mextabotrpic responses to ACh and 5-HT of Xenopus oocytes injetcted with rat brain mRNA are transduced by different G-protein subtypes" FEBS Lett.299. 179-182 (1992)

Shuji Kaneko：“注射了大鼠脑 mRNA 的爪蟾卵母细胞对 ACh 和 5-HT 的 Mextabotrpic 反应是由不同的 G 蛋白亚型转导的”FEBS Lett.299。

Shuji Kaneko, Junichi Yuasa, Hideki Takahashi and Masamichi Satoh: "Expression of opioid receptors in Xenopus oocytes injected with rat brain mRNA" Soc. Neurosci. Abst.18. 1169 (1992)

Shuji Kaneko、Junichi Yuasa、Hideki Takahashi 和 Masamichi Satoh：“注射大鼠脑 mRNA 的爪蟾卵母细胞中阿片受体的表达”Soc。