アフリカツメガエル卵母細胞発現系を用いたオピオイド受容体の脱感作機構の解析

利用非洲爪蟾卵母细胞表达系统分析阿片受体脱敏机制

• 批准号: 06770070
• 负责人: 宮前 丈明
• 金额: $ 0.58万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06770070/
• 关键词: アフリカツメガエル; オピオイド受容体; GTP結合蛋白質; 脱感作; リン酸化; プロテインキナーゼC

本研究者は既に、アフリカツメガエル卵母細胞発現系を用いてクローン化されたオピオイドδ受容体と外来のGTP結合蛋白質(G-蛋白)αサブユニットを同時に発現させた、いわゆる「機能的受容体-G-蛋白再構成モデル」を作成し、NG108-15細胞由来のオピオイドδ受容体(DOR-1)がGi-蛋白(Gilα)と共役してホスホリパーゼC活性化にともなうCa^<2+>依存性Cl-電流(Icl^-(Ca^<2+>))を誘発することを報告した(Miyamae T et al. 1993,FEBS Lett.333,311-314)。本年度はこのモデル系を用いてオピオイドδ受容体応答の脱感受性機構の解析を進めた。その結果、(1)同受容体応答は、高濃度(μMオーダー)のアゴニストの頻回適用によってアゴニスト依存性の脱感受性反応を生じること、(2)この脱感受性反応はcalphostin CなどのプロテインキナーゼC(PKC)阻害薬やPKC阻害ペプチドによって特異的に抑制されること、一方cAMP依存性キナーゼ(PKA)、チロシンキナーゼ、及びカルモジュリン依存性キナーゼ(CaMKII)などに特異的な阻害薬ないし阻害ペプチドは無作用であること、(3)PKC阻害薬或いは阻害ペプチドは、G-蛋白を直接活性化する作用を持つAlF_4-の適用によって惹起されるIcl^-(Ca^<2+>)に対し無作用であること、などの知見が集積された。これらの結果から、オピオイドδ受容体応答のアゴニスト依存性の脱感受性反応は、受容体以後ではなく受容体自身がPKCによってリン酸化されることにより生じることが強く示唆された(Ueda H,Miyamae T et al. 1994,Regulatory Peptides 54,305-306)。従って本年度は、同受容体のシグナル伝達制御機構解明への端緒とも言うべき重要な一知見を得ることができた。

In this study, both the oocyte receptor and the receptor of the oocyte were used to induce the expression of the foreign GTP binding protein (G-protein) alpha receptor protein (G-protein). At the same time, the receptor-G-protein of the function of the receptor-G-protein was reproduced to form the receptor. The origin of NG108-15 cells is that the receptor delta receptor (DOR-1), the Gi- protein (Gil α), the co-active protein (Gil α), the activated protein C (C-activating protein), the δ-receptor (DOR-1) and the protein (Gil α). (2) dependent Cl- current (ICL ^-(CA ^ & lt;2+>)) is responsible for reporting and reporting (Miyamae T et al.) 1993 Febs Lett.333311-314) This year, the system is based on the results of the analysis of the desensitization mechanism. The results were as follows: (1) homoreceptive response, high concentration (μ M), high concentration (μ M), high temperature (μ M), (1) homoreceptive response, (1) high concentration (μ M), (1) homoreceptive response, (1) homoreceptor response, (1) high concentration (μ M One side camp dependent drugs (PKA), one side camp dependent drugs (PKA), one side camp dependent drugs (CaMKII), one side camp dependent drugs (lt), and one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), one side camp dependent drugs (camp dependent drugs), and one side camp dependent drugs (lt). (2) to make sure that you have the right to know what you want to know. The results showed that the receptor responded to the dependence, desensitization and anti-receptivity, and the acceptor was followed by the recipient itself. The PKC was acidified and acidified. The recipient strongly indicated that it was instigated by Ueda. 1994 54305 Regulatory Peptides. In the current year, the same recipient system has made it clear that it is very important to know that it is important to know that it is important to make a decision.

项目成果

Miyamae,T: "Involvement of phosphorylation mechanisms in delta opioid receptor-mediated phospholipase C activation in Xenopus oocytes." Neurosci.Res.suppl.19. S58- (1994)

Miyamae,T：“磷酸化机制参与非洲爪蟾卵母细胞中 δ 阿片受体介导的磷脂酶 C 激活。”