Cloning, gene expression and signal transduction of opioid kappa-receptor

阿片κ受体的克隆、基因表达及信号转导

• 批准号: 05454577
• 负责人: UEDA Hiroshi
• 金额: $ 4.48万
• 依托单位: Yokohama City Univ.
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05454577/
• 关键词: Opioid receptor; Signal transduction; Xenopus Laevis; Oocytes; Cloning; G-protein; 〓イノフィンACセプター; オピオイドレセプター; 情報伝達; 母細胞; 発現クローニング

Here we report the evidence for a novel type of opioid kappa-receptor inhibiting G-protein activity in the guinea pig cerebellum. In guinea pig cerebellar membranes, kappa-agonists inhibit the high-affinity GTPase activity in concentrations lower than those for stimulation of this activity, possibly through known 7-transmembrane type of opioid kappa-receptor. The inhibitory activity was found to be attributed to the direct inhibition of GTP-GDP exchange activity on Gil reconstituted into membranes. Taken into consideration the finding that kappa-receptor agonists inhibit phospholipase C through an inhibition of Gil activity, we attempted to clone this metabostatic receptor which inhibits currents mediated through Gil and phospholipase C in Xenopus oocytes. In Xenopus oocytes coinjected with RNAs of metabotropic glutamate receptor (or muscarinic M2 receptor) and Gila, the kappa-receptor agonists inhibits these metabotropic receptor-mediated currents. These inhibitory effects were completely antagonized by norBNI,an opioid kappa-receptor antagonist. Using conventional strategies, we cloned a novel type of metabostatic kappa-receptor which has no apparent transmembrane domain.

在这里，我们报告了一种新型阿片κ受体抑制豚鼠小脑G蛋白活性的证据。在豚鼠小脑膜中，κ-激动剂在低于刺激该活性的浓度下抑制高亲和力GTdR活性，可能通过已知的7-跨膜型阿片κ-受体。发现抑制活性归因于直接抑制重组到膜中的Gil的GTP-GDP交换活性。考虑到这一发现，κ受体激动剂抑制磷脂酶C通过抑制Gil活性，我们试图克隆这种代谢抑制受体，抑制电流介导的Gil和磷脂酶C在非洲爪蟾卵母细胞。在非洲爪蟾卵母细胞共注射RNA的代谢型谷氨酸受体（或毒蕈碱M2受体）和Gila，κ受体激动剂抑制这些代谢型受体介导的电流。这些抑制作用被阿片κ受体拮抗剂norBNI完全拮抗。利用传统的策略，我们克隆了一种新型的代谢抑制κ受体，它没有明显的跨膜结构域。

项目成果

Ueda at al.: "Protein kinase inhibitor protentiatcs opitid α-receptor-currents in Xenopus oocytes" Neuro Report. 5. 1985-1988 (1994)

Ueda 等人：“爪蟾卵母细胞中的蛋白激酶抑制剂 protentiatcs optid α-受体电流”《神经报告》5. 1985-1988 (1994)。

Miyamae, T., Fukushima, N., Misu, Y.and UEDA,H: "delta Opioid receptor mediates phospholipase C activation via Gi in Xenopus oocytes." Febs Lett. 333. 311-314 (1993)

Miyamae, T.、Fukushima, N.、Misu, Y. 和 UEDA,H：“δ 阿片受体通过爪蟾卵母细胞中的 Gi 介导磷脂酶 C 激活。”

Ueda et al: "Supersensitigation of neurchemical responses by L-DOPA and dopamine receptor agorirots in the striatum of experimental Parkorseris disease model rats" Biomedicine and Pharmacethrepy. 49. 169-177 (1995)

Ueda 等人：“实验性 Parkorseris 疾病模型大鼠纹状体中 L-DOPA 和多巴胺受体 agorirots 对神经化学反应的超敏化”生物医学和药剂学。

UEDA,H., Miyamae, T., Fukushima, N.and Misu, Y: "Protein kinase inhibitor poten tiates opioid delta-receptor-currents in X enopus oocytes." NeuroReport. 5. 1985-1988 (1994)

UEDA,H.、Miyamae, T.、Fukushima, N. 和 Misu, Y：“蛋白激酶抑制剂增强 X enopus 卵母细胞中的阿片类 δ 受体电流。”

Fukushima, N., UEDA,H., Hayashi, C., Katayama, T., Miyamae, T.and Misu, Y: "Species and age-dependent differences of functional coupling between opioid delta-receptor and G-proteins and possible involvement of protein kinase C in striatal membranes." Neur

Fukushima, N.、UEDA,H.、Hayashi, C.、Katayama, T.、Miyamae, T. 和 Misu, Y：“阿片类药物 δ 受体和 G 蛋白之间功能耦合的物种和年龄依赖性差异以及可能的