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Functional Analysis of small G protein during cardiac early morphogenesis

小G蛋白在心脏早期形态发生过程中的功能分析

基本信息

批准号：
12670769
负责人：
SHIRAISHI Isao
金额：
$ 2.24万
依托单位：
Kyoto Prefectural University of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670769/
关键词：
small G protein Rho FAK cardiac morphogenesis 先天性心奇形心ループ低分子G蛋白質筋原線維

项目摘要

To understand the mechanism of normal myofibrillogenesis in vivo, we have reported 3D observation of myofibril formation, cell-cell and cell-matrix adhesions, and tyrosine phosphorylation during looping of whole-mounted chicken embryonic heart. We found that formation and arrangement of myofibrils develops in close association with changing cell-cell and cell-matrix adhesions. In this study, we performed 3D observation of focal adhesion kinase (FAK) and paxillin immunolocalization before and during looping (4- to 15-somite stages) in the same manner. Before myofibrillogenesis (4 to 7-somite stages), faint and particulate pattern of FAK and paxillin was randomly expressed at the bottom of the inner myocardial cell layer facing the cardiac jelly. During initial stages of myofibrillogenesis (8 to 9-somite stages), fibrillar pattern of FAK and paxillin appeared at termini of stress fiber-like thick actin bundles which were recognized as an initial stage of myofibrillogensis. During active stages of myofibrillogenesis (10-13-somite stages), paxillin was also localized at Z-bands of immature striated myofibrils, while FAK was not co-localized with Z-bands. At the 14-15 somite stages or later, the expression of FAK and paxillin diminished especially at sites of mature striated myofibrils. These results suggests that paxillin is involved in the initial arrangement and sarcomere formation of cardiac myofibrils, while FAK is involved in the initial arrangement of myofibrils.

为了了解体内正常肌原纤维形成的机制，我们报道了三维观察肌原纤维的形成，细胞-细胞和细胞-基质粘附，和酪氨酸磷酸化在成环的鸡胚心脏。我们发现，肌原纤维的形成和排列与细胞-细胞和细胞-基质粘附的变化密切相关。在这项研究中，我们进行了3D观察粘着斑激酶（FAK）和桩蛋白免疫定位之前和期间成环（4至15体节阶段）以同样的方式。在肌原纤维形成（4 - 7体节阶段）之前，FAK和桩蛋白的微弱和颗粒图案随机表达于面向心胶质的内心肌细胞层的底部。在肌原纤维发生的初始阶段（8 - 9体节阶段），FAK和桩蛋白的纤维模式出现在应力纤维样的粗肌动蛋白束的末端，这被认为是肌原纤维发生的初始阶段。在肌原纤维发生的活跃阶段（10-13体节阶段），桩蛋白也定位于未成熟横纹肌原纤维的Z带，而FAK不与Z带共定位。在14-15体节期或以后，FAK和桩蛋白的表达减少，尤其是在成熟的横纹肌原纤维部位。这些结果表明，桩蛋白参与了心肌肌原纤维的初始排列和肌节的形成，而FAK参与了肌原纤维的初始排列。