Molecular and functional analysis of ATP-sensitive K^+ channel on the nuclear envelope

核膜上 ATP 敏感 K^ 通道的分子和功能分析

• 批准号: 18590232
• 负责人: NAKAYA Haruaki
• 金额: $ 2.57万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590232/
• 关键词: nuclear envelop; ATP-sensitive K^+ channel; sufonylurea drug; K^+ channel opener; Heart; 薬理学; 生理学; 細胞・組織

This study was undertaken to test whether ATP-sensitive K^+ (K_<ATP> ) channel exists on the nuclear envelope of cardiac cells and to determine the functional molecules and physiological roles of K_<ATP> channel on the nuclear envelope. Using binding assay of glibenclamide-BODIPY-FL to sulfonylurea receptor we tested the existence of K_<ATP> channel on the nuclear envelope. We could observe the binding of glibenclamide-BODIPY-FL colocalized with Hoechst, a nuclear marker, using confocal microscopy, suggesting the existence of sufonylurea receptor on the nuclear envelope of cardiomyocytes. The single channel activity was recorded from the nuclear envelope of isolated nuclei using patch clamp techniques. Although several kinds of single channel activity could be recorded from the nuclear envelope, the channel activity was not influenced by glibenclamide, tolbutamide, or pinacidil. Nuclear Ca^<2+> signal was measured using confocal microscopy and the Ca^<2+> sensitive fluorescence dye Fluo-3/AM. Neither glibenclamide nor tolbutamide affected Ca^<2+> concentration in the nuclear envelope. Thus, even if K_<ATP> channel exists on the nuclear envelope of cardiac cells, the density or physiological significance of the channel may not be so large.

本研究旨在检测心肌细胞核膜上是否存在ATP敏感的K^+（K_<ATP>）通道，并探讨核膜上K_<ATP>通道的功能分子和生理作用。使用格列本脲-BODIPY-FL与磺酰脲受体的结合测定，我们测试了核膜上K_<ATP>通道的存在。我们可以使用共聚焦显微镜观察到格列本脲-BODIPY-FL与核标记物Hoechst共定位的结合，表明心肌细胞核膜上存在磺脲类受体。使用膜片钳技术从分离的细胞核的核膜记录单通道活性。尽管可以从核膜记录到多种单通道活性，但通道活性不受格列本脲、甲苯磺丁脲或吡那地尔的影响。使用共聚焦显微镜和Ca 2+ 敏感荧光染料Fluo-3/AM测量核Ca 2+ 信号。格列本脲和甲苯磺丁脲都不影响核膜中的Ca 2+ 浓度。因此，即使心肌细胞的核膜上存在K_<ATP>通道，该通道的密度或生理意义也可能不会那么大。

项目成果

心筋細胞K^+チャンネルおよびミトコンドリアに対するベプリジルの作用

苯普地尔对心肌细胞K^+通道和线粒体的影响

• 发表时间: 2007
• 作者: 中谷晴昭

Role of ATP-sensitive K^+ channels in cardioprotection

ATP 敏感 K^ 通道在心脏保护中的作用

• 发表时间: 2006
• 作者: 中谷晴昭

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• 发表时间: 2006
• 作者: Yamashita Y

Evaluation of arrhythmogenic properties of new drugs

新药致心律失常特性评价

• 发表时间: 2008
• 作者: Nakaya H.

抗心房細動薬の展望-新しい標的を求めて-

抗心房颤动药物的前景 - 寻找新靶点 -

• 发表时间: 2007
• 作者: 中谷 晴昭