Mechanism of the cytoskeletal regulation of store-operated Ca^<2+> entry

钙池操纵的Ca^<2>进入的细胞骨架调节机制

• 批准号: 16590165
• 负责人: OMATSU Mariko
• 金额: $ 1.98万
• 依托单位: Shiga University of Medical Science
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590165/
• 关键词: store-operated Ca^<2+> entry; P2 receptor; ATP; intracellular Ca^<2+>; cytoskeleton; actin; endoplasmic reticulum; brown adipocyte; 容量生カルシウム流入

Stimulation of P2 receptors with micromolar concentration of ATP not only evokes a transient increase in intracellular free Ca^<2+> concentration ([Ca^<2+>]_i), primarily due to release of Ca^<2+> from intracellular stores ; such stimulation also triggers almost complete suppression of thapsigargin-evoked sustained [Ca^<2+>]_i elevation mediated through a store-operated Ca^<2+> entry pathway in rat brown adipocytes. We investigated the role of cytoskeletal actin in the inhibitory effect which extracellular ATP has on store-operated Ca^<2+> entry, using fura-2 fluorescence for continuous measurement of [Ca^<2+>]_i_, and using Alexa fluor 488-phalloidin staining of actin. Disassembly of actin networks by cytochalasin D (1 μM) or latrunculin A (3 μM) prevented the inhibitory effect of ATP (10 μM) on the thapsigargin (100 μM)-evoked store-operated Ca^<2+>entry, without changing the effect of ATP in raising [Ca^<2+>]_i. In normal cells, bath application of ATP induced a transient [Ca^<2+>]_i elevation consisting of a rapid increase (the rising phase) and the subsequent decrease (the declining phase) to a lower steady level despite the continued presence of the agonist. Disruption of actin assemblies did not significantly affect the rising phase, but prevented the declining phase. Cells incubated with 10 μM ATP for 4 min demonstrated marked accumulations of actin filaments at the cell periphery, showing protrusions at the cell surface ; this actin-assembly process is mediated through P2 receptors. In cells treated with cytochalasin D or latrunculin A, extracellular ATP did not induce actin redistribution. These results suggest the actin reorganization plays a role in ATP-induced inhibition of store-operated Ca^<2+> entry in rat brown adipocytes.

用微摩尔浓度的 ATP 刺激 P2 受体不仅引起细胞内游离 Ca^2+ 浓度 ([Ca^2+]_i) 的短暂增加，这主要是由于细胞内储存的 Ca^2+ 的释放；这种刺激还触发了对毒胡萝卜素引起的持续[Ca ^ 2+ ]_i升高的几乎完全抑制，该持续[Ca ^ 2+ ]_i升高是通过大鼠棕色脂肪细胞中钙库操纵的Ca ^ 2+ 进入途径介导的。我们研究了细胞骨架肌动蛋白在细胞外 ATP 对钙池操纵的 Ca^2+ 进入的抑制作用中的作用，使用 fura-2 荧光连续测量 [Ca^2+]_i_，并使用 Alexa flor 488-鬼笔环肽染色肌动蛋白。细胞松弛素 D (1 μM) 或 latrunculin A (3 μM) 分解肌动蛋白网络可防止 ATP (10 μM) 对毒胡萝卜素 (100 μM) 诱发的钙池操作的 Ca^2+ 进入的抑制作用，而不改变 ATP 在提高 [Ca^2+]_i 中的作用。在正常细胞中，尽管激动剂持续存在，但ATP的浴应用诱导短暂的[Ca 2+ ]_i升高，其包括快速增加(上升阶段)和随后降低(下降阶段)至较低的稳定水平。肌动蛋白组装的破坏并没有显着影响上升期，但阻止了下降期。细胞与10μM ATP孵育4分钟，细胞外围肌动蛋白丝明显聚集，细胞表面出现突起；该肌动蛋白组装过程是通过 P2 受体介导的。在用细胞松弛素 D 或 latrunculin A 处理的细胞中，细胞外 ATP 不会诱导肌动蛋白重新分布。这些结果表明肌动蛋白重组在大鼠棕色脂肪细胞中ATP诱导的钙池操纵的Ca 2+ 进入抑制中发挥作用。

项目成果

Cytosolic Ca2+ under high glucose with suppressed Na+/K+ pump activity in rat sensory neurons

• DOI: 10.1097/00001756-200401190-00038
• 发表时间: 2004-01
• 期刊: NeuroReport
• 影响因子: 1.7
• 作者: M. Sanada;H. Matsuura;M. Omatsu-Kanbe;K. Sango;A. Kashiwagi;H. Yasuda

Angiotensin II potentiates the slow component of delayed rectifier K^+ current via AT_1 receptor in guinea-pig atrial myocytes.

血管紧张素 II 通过豚鼠心房肌细胞中的 AT_1 受体增强延迟整流 Kk电流的慢分量。

• 发表时间: 2006
• 期刊: Circulation 113
• 作者: Kawajiri;M.;Okano;Y.;Kuno;M.;Hase;Y.;Inada;H.;Tashiro;F.;Miyazaki;J.;Yamono;Y.;Zankov D.P.
• 通讯作者: Zankov D.P.

Monkey embryonic stem cells differentiate into adipocytes in vitro

• DOI: 10.1089/clo.2006.8.3
• 发表时间: 2006-03-01
• 期刊: CLONING AND STEM CELLS
• 作者: Yamashita, A;Takada, T;Torii, R
• 通讯作者: Torii, R

Identification of endogenous surrogate ligands for human P2Y receptors through an in silico search

• DOI: 10.1254/jphs.95.81
• 发表时间: 2004-05-01
• 期刊: JOURNAL OF PHARMACOLOGICAL SCIENCES
• 影响因子: 3.5
• 作者: Hiramoto, T;Nonaka, Y;Fujita, N
• 通讯作者: Fujita, N

Actin filaments play permissive role in the inhibition of store-operated Ca^<2+> entry by extracellular ATP in rat brown adipocytes.

在大鼠棕色脂肪细胞中，肌动蛋白丝在抑制胞外ATP通过钙池操纵的Ca 2+ 进入中发挥许可作用。

• 发表时间: 2004
• 期刊: Biochem.J. 381
• 作者: Omatsu-Kanbe;M.;Shibata;M.;Yamamoto;T.;Isono;T.;Matsuura,H.
• 通讯作者: Matsuura,H.