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，并确定K_<ATP>通道在核膜上的功能分子和生理作用。采用格列本脲- bodipy - fl与磺酰脲受体的结合实验，验证了核膜上是否存在K_<ATP>通道。我们利用共聚焦显微镜观察到glibenclamide-BODIPY-FL与细胞核标记物Hoechst共定位的结合，提示心肌细胞核膜上存在磺酰脲受体。用膜片钳技术记录了离体核包膜的单通道活动。虽然可以从核膜上记录到几种单通道活性，但通道活性不受格列本脲、甲苯丁酰胺或品酸酯的影响。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。格列本脲和甲苯丁酰胺均不影响核膜Ca^<2+>浓度。因此，即使心脏细胞核膜上存在K_<ATP>通道，该通道的密度或生理意义也可能没有那么大。

项目成果

Electrophysiological roles of ATP-sensitive K^+ channels in mouse sinoatrial node

小鼠窦房结中 ATP 敏感 K^ 通道的电生理作用

• 发表时间: 2007
• 作者: 土屋 浩一郎;Nishida H.;Fujita T.;Yasuda N.;Yasuda N.;Nishida A.;中谷 晴昭;中谷 晴昭;中谷 晴昭;中谷晴昭;Yamashita Y;Yamashita Y.;Sato T.;中谷晴昭;Nakaya H.;Fukuzaki K.
• 通讯作者: Fukuzaki K.

心房細動薬物治療の標的分子

房颤药物治疗的靶分子

• 发表时间: 2007
• 期刊: PM基礎・治療 27
• 作者: Nishida A.;Reien Y.;Ogura T.;Uemura H.;Tamagawa M.;Yabana H.;Nakaya H.;中谷晴昭;中谷晴昭;Nishida A.;中谷 晴昭;中谷 晴昭;中谷 晴昭;中谷 晴昭
• 通讯作者: 中谷 晴昭

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
• 期刊: 心電図 27・1
• 作者: 土屋 浩一郎;Nishida H.;Fujita T.;Yasuda N.;Yasuda N.;Nishida A.;中谷 晴昭;中谷 晴昭;中谷 晴昭;中谷晴昭
• 通讯作者: 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: 土屋 浩一郎;Nishida H.;Fujita T.;Yasuda N.;Yasuda N.;Nishida A.;中谷 晴昭;中谷 晴昭;中谷 晴昭;中谷晴昭;Yamashita Y;Yamashita Y.;Sato T.;中谷晴昭;Nakaya H.;Fukuzaki K.;中谷晴昭
• 通讯作者: 中谷晴昭

Effects of azimilide on the muscarinic acetylcholine receptoroperated K^+ current and experimental atrial fibrillation in guinea-pig hearts

阿齐利特对豚鼠心脏毒蕈碱乙酰胆碱受体操纵的K^电流和实验性心房颤动的影响

• 发表时间: 2007
• 期刊: J. Pharmacol. Sci. 105
• 作者: Nishida A.;Reien Y.;Ogura T.;Uemura H.;Tamagawa M.;Yabana H.;Nakaya H.
• 通讯作者: Nakaya H.