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.

本研究的目的是探讨胃肠道内神经元容量性钙离子内流的调控机制。出于这些目的，我们分析了容量性Ca内流的特性、其与细胞内信使和推定通道分子的调节机制。1)我们建立了单个培养的胃肠内源性神经元钙信号可视化的实验系统。2)ATP引起大鼠胃肠神经元内钙增加，其作用机制包括胞内钙释放、钙内流和P2X2通道激活。3)缓激肽通过肠神经胶质细胞分泌的PGE2激活胃肠内源性神经元上的B2受体而引起Ca的升高。4)在PC12细胞中表达的克隆TRPC 5通道作为与GTP结合蛋白受体偶联的Ca进入通道发挥作用。5)克隆TRPV1的互补DNA，并在HEK293细胞中异源表达。我们发现，TRPV1通道功能的多模态受体通道与高钙渗透性。这些结果表明，容量性Ca内流通道在胃肠内源性神经元的功能性Ca内流通路中起作用，该通路受多种内源性因子如ATP和缓激肽的调节。克隆的TRPC 5和TRPV 1构成调节细胞内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

Zinc modulates primary afferent fiber-evoked responses of ventral roots in neonatal rat spinal cord in vitro

锌在体外调节新生大鼠脊髓腹侧根的初级传入纤维诱发反应

• 发表时间: 2006
• 期刊: Neuroscience 138
• 作者: Ito Y;Katsura K;Maruyama K;Taji T;Kobayashi M;Seki M;Shinozaki K;Yamaguchi-Shinozaki K;Ken-ichi OTSUGURO
• 通讯作者: Ken-ichi OTSUGURO

Redox regulation of PI3K/Akt and p53 in bovine aortic endothelial cells exposed to hydrogen peroxide

• DOI: 10.1089/152308603770380016
• 发表时间: 2003-12-01
• 期刊: ANTIOXIDANTS & REDOX SIGNALING
• 影响因子: 6.6
• 作者: Niwa, K;Inanami, O;Kuwabara, M
• 通讯作者: Kuwabara, M

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”

P2X2 receptors are essential for [Ca2+]i increases in response to ATP in cultured rat myenteric neurons.

• DOI: 10.1152/ajpgi.00017.2005
• 发表时间: 2005-05
• 期刊: American journal of physiology. Gastrointestinal and liver physiology
• 作者: T. Ohta;Akane Kubota;M. Murakami;K. Otsuguro;S. Ito