Studies on the regulation of myocardial SR calcium release channel under in situ environment

原位环境下心肌SR钙释放通道调控研究

• 批准号: 11670105
• 负责人: TANAKA Hikaru
• 金额: $ 2.11万
• 依托单位: TOHO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670105/
• 关键词: mouse myocardium; sarcoplasmic reticulum; calcium ion; cameleon; adenovirus vector; ryanodine receptor; リアノジン受容体; Ca^<2+>スパーク; fluo-3; 共焦点顕微鏡; T管; ミトコンドリア

cDNAs for Yellow Cameleons with targeting sequences towards the mitochondria and endoplasmic reticulum were packaged into an inproved adenoviral vector and were introduced to primary cultured cardiomyocytes and hepatocytes. The targetting of the probes to organella were confirmed by comparing the fluorescence with those of TMRE etc. Difference in excitation-contraction mechanisms between the atria and ventricle were examined with rapid scanning confocal microscopy. A propagation of CICR mechanisms was involved in normal EC coupling in atrial myocytes but not in ventricular myocytes. This difference was due to the absence of T-tubules in atrial myocytes rather than difference in the properties of individual release unitsand might explain some of the pharmacological difference between the atria and ventricle. We examined the role of three membrane proteins in triggering calcium release from the ryanodine receptor channel. In both atrial and ventricular cardiomyocytes, L-type, but not T-type calcium channels were involved in triggering calcium release. The sodium-calcium exchanger, when activated by factors such as alpha adrenergic stimulation, was found to reduce calcium release from the sarcoplasmic reticulum indirectly through decrease in calcium load.

将靶向线粒体和内质网的靶向序列的黄色粉红色包装到侵略性腺病毒载体中，并被引入原代培养的心肌细胞和肝细胞中。通过将荧光与TMRE的荧光进行比较，可以通过快速扫描共聚焦显微镜检查中心房和心室之间的激发反应机制的差异来确认探针对有机疾病的靶向。 CICR机制的传播参与了心肌细胞中的正常EC偶联，但在心室心肌细胞中不参与。这种差异是由于心肌细胞中缺乏T管，而不是单个释放单位的性质差异，并且可能解释了心房和心室之间的某些药理差异。我们研究了三种膜蛋白在触发Ryanodine受体通道释放钙释放中的作用。在心房和心室心肌细胞中，L型，但不参与T型钙通道触发钙释放。当被诸如α肾上腺素能刺激之类的因子激活时，发现钠钙的交换剂可通过钙负荷下降间接地从肌质网中释放钙释放。

项目成果

Masumiya H., Kase J., Tanaka Y., Tanaka H., Shigenobu K.: "Effects of mibefradil, a selective T-type Ca^<2+> channel antagonist on sino-atrial node and ventricular myocardia."Res.Comm.Mol.Path.Pharmacol.. 104. 321-329 (1999)

Masumiya H.、Kase J.、Tanaka Y.、Tanaka H.、Shigenobu K.：“米贝拉地尔（一种选择性 T 型 Ca^<2> 通道拮抗剂）对窦房结和心室心肌的影响。”Res.Comm

Nishimaru K., Tanaka Y, Tanaka H., Shisenobu K.: "α-adrenoceptor stimulation-mediated negative inotropism and enhancement of Na^+-Ca^<2+> exchange in mouse ventricle"Am.J.Physiol. 280. H132-H141 (2001)

Nishimaru K.、Tanaka Y、Tanaka H.、Shisenobu K.：“α-肾上腺素受体刺激介导的负性肌力作用和小鼠心室 Na^+-Ca^<2+> 交换的增强”Am.J.Physiol。 H132-H141 (2001)

Nishimaru K. et al.,: "d-adrenoceptor stimulation-induced negative inotropism and enhancement of Na^+-Ca^<2+> exchangers in mouse ventricle"Am.J.Physiol.. 280. H132-H141 (2001)

Nishimaru K.等人，：“小鼠心室中d-肾上腺素受体刺激诱导的负性肌力作用和Na ^ -Ca ^ 2 >交换器的增强”Am.J.Physiol..280.H132-H141(2001)

Masumiya H et al.: "Effects of mibefradil, a selective T-type Ca^<2+> channel antagonist on SA node and ventricular myocardia."Res.Comm.Mol.Path.Pharmacol.. 104. 321-329 (1999)

Masumiya H 等人：“米贝拉地尔，一种选择性 T 型 Ca^2 通道拮抗剂对 SA 结和心室心肌的影响。”Res.Comm.Mol.Path.Pharmacol.. 104. 321-329 (1999)

T.Sekine,H.Kusano,K.Nishimaru,Y.Tanaka,H.Tanaka,K.Shigenobu: "Developmental Conversion of inotropism by endothelin 1 and angiotensinII from posirive to negative in mice"European Journal of Pharmacology. 374. 411-415 (1999)

T.Sekine、H.Kusano、K.Nishimaru、Y.Tanaka、H.Tanaka、K.Shigenobu：“小鼠中内皮素 1 和血管紧张素 II 的正性肌力从阳性到阴性的发育性转化”欧洲药理学杂志。