Role of store-operated Ca entry mechanism in signal transduction in gastrointestinal intrinsic neurons

钙池操纵的钙进入机制在胃肠固有神经元信号转导中的作用

• 批准号: 15380200
• 负责人: OHTA Toshio
• 金额: $ 8.77万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380200/
• 关键词: intracellular calcium; bradykinin; ATP; glia; TRP; Ca imaging; intracellular Ca store; channel; ブラジキンン; プロスタグランジン; シクロオキシゲナーゼ; 細胞内Ca; パッチクランプ; 蛍光画像解析; カルシウムチャンネル; アデニンヌクレオチド

The aim of this project was to identify the regulatory mechanisms of capacitative Ca entry in gastrointestinal intrinsic neurons. For these purposes, we analyzed the properties of capacitative Ca entry, its regulatory mechanisms with intracellular messengers and putative channel molecules. 1) We established experimental system for visualization of Ca signaling in single cultured gastrointestinal intrinsic neurons. 2) ATP elicited Ca increases through release of Ca from intracellular stores, the activation of capacitative Ca entry and P2X2 channels in rat cultured gastrointestinal intrinsic neurons. 3) Bradykinin evoked the elevation of Ca through B2 receptor activation on gastrointestinal intrinsic neurons via PGE2 secreted from enteric glial cells. 4) Cloned TRPC5 channels expressed in PC12 cells functioned as Ca entry channels coupled with GTP-binding protein receptors. 5) Complimentary DNA for TRPV1 was cloned and was hetelogeously expressed in HEK293 cells. We revealed that TRPV1 channels functioned as polymodal receptor channels with high Ca permeability. These results indicate that capacitative Ca entry channels play a role for functional Ca entry pathways in gastrointestinal intrinsic neurons, which are regulated by various intrinsic factors such as ATP and bradykinin. Cloned TRPC5 and TRPV1 constitute Ca entry channels for regulation of intracellular Ca signallings.

该项目的目的是确定胃肠固有神经元电容性 Ca 进入的调节机制。为此，我们分析了电容性 Ca2+ 进入的特性、其与细胞内信使和假定通道分子的调节机制。 1）我们建立了单个培养的胃肠固有神经元中Ca信号传导可视化的实验系统。 2) ATP 通过细胞内储存的 Ca 释放、大鼠培养的胃肠固有神经元中电容性 Ca 进入和 P2X2 通道的激活而引起 Ca 增加。 3) 缓激肽通过肠道神经胶质细胞分泌的 PGE2 激活胃肠固有神经元上的 B2 受体，从而引起 Ca 升高。 4)在PC12细胞中表达的克隆TRPC5通道充当与GTP结合蛋白受体偶联的Ca进入通道。 5)TRPV1的互补DNA被克隆并在HEK293细胞中异源表达。我们发现 TRPV1 通道充当具有高 Ca 渗透性的多模式受体通道。这些结果表明，电容性 Ca2+ 进入通道在胃肠固有神经元的功能性 Ca2+ 进入途径中发挥作用，该途径受到 ATP 和缓激肽等各种内在因子的调节。克隆的 TRPC5 和 TRPV1 构成了调节细胞内 Ca 信号传导的 Ca 进入通道。

项目成果

P2X_2 receptors are essential for [Ca^<2+>]_1 increases in response to ATP in cultured rat myenteric neurons.

P2X_2 受体对于培养的大鼠肌间神经元中响应 ATP 的 [Ca^2>]_1 增加至关重要。

• 发表时间: 2005
• 期刊: American Journal of Physiology 298
• 作者: Otsuguro K.;Ohta T.;Ito S.;Ohta T
• 通讯作者: Ohta T

Ohta T., Morishita M., Mori Y., Ito S.: "Ca^<2+> store-independent augmentation of [Ca^<2+>]_i responses to G-protein coupled receptor activation in recombinantly TRPC5-epressed rat pheochromocytoma (PC12) cells"Neurosci.Lett.. 358. 161-164 (2004)

Ohta T.、Morishita M.、Mori Y.、Ito S.：“在重组 TRPC5 抑制的大鼠嗜铬细胞瘤中，[Ca^2>]_i 对 G 蛋白偶联受体激活的反应不依赖于 Ca^2> 储存而增强”

Molecular cloning, functional characterization of the porcine transient receptor potential V1 (pTRPV1) and pharmacological comparison with endogenous pTRPV.

猪瞬时受体电位 V1 (pTRPV1) 的分子克隆、功能表征以及与内源性 pTRPV 的药理学比较。

• 发表时间: 2005
• 期刊: Biochemical Pharmacology 71
• 作者: Fujisawa M;Kiyosue M;Hori M;Ozaki H;Ohta T;Murakami M;Ohta T;Takahashi E;Otsuguro K;Otsuguro K;Otsuguro K;Ohta T;Ohta T
• 通讯作者: Ohta T