Wolf W P, Spicher K, Haase H, Schulze W
Max-Delbrück-Centrum für Molekulare Medizin, Berlin, Germany.
J Mol Cell Cardiol. 1998 Jun;30(6):1149-62. doi: 10.1006/jmcc.1998.0679.
The cellular and sub-cellular localization of the G-protein subunit, G(o) alpha, in rat heart was determined by immunofluorescence and immunoelectron microscopy. Using antibodies directed against purified G(o) alpha and an antiserum raised against the C-terminal decapeptide of G(o) alpha, strong immunoreactivity was found in the conducting system of the heart, neurons, and atrial cardiomyocytes. Labeling of ventricles was weak compared to that of atria. In neurons, immunoelectron microscopy revealed G(o) alpha was localized along the inner surface of axolemma and on axoplasmal vesicles. G(o) alpha immunoreactivity in atrial and ventricular myocytes was not restricted to sarcolemma, but was also found on sub-sarcolemmal vesicles with characteristic caveolar morphology. At the level of intercalated discs, labeling was spread over the periphery of intercalated discs avoiding its membrane structures. Additionally, in atrial endocrine cardiomyocytes, approximately 60% of secretory granules revealed G(o) alpha-labeling on the cytoplasmic surface. A small number of these granules stood out due to particularly intense labeling. The observation that these granules were found most-frequently in sub-sarcolemmal areas suggests that they may be mature granules undergoing exocytosis. Therefore, G(o) alpha found on secretory granules of endocrine cardiomyocytes may have a function in regulated exocytosis of cardiac hormones. Sarcolemmal localization of G(o) alpha in atrial and ventricular cardiomyocytes supports the role of G(o) alpha in transmembrane signal transduction. Furthermore, caveolar localization of G(o) alpha may provide a compartmental basis for integrating G(o)-mediated signaling events.
通过免疫荧光和免疫电子显微镜确定了G蛋白亚基G(o)α在大鼠心脏中的细胞和亚细胞定位。使用针对纯化的G(o)α的抗体和针对G(o)α C末端十肽产生的抗血清,在心脏传导系统、神经元和心房心肌细胞中发现了强免疫反应性。与心房相比,心室的标记较弱。在神经元中,免疫电子显微镜显示G(o)α定位于轴膜内表面和轴浆小泡上。心房和心室肌细胞中的G(o)α免疫反应性不仅限于肌膜,还存在于具有特征性小窝形态的肌膜下小泡上。在闰盘水平,标记分布在闰盘周边,避开其膜结构。此外,在心房内分泌心肌细胞中,约60%的分泌颗粒在细胞质表面显示G(o)α标记。少数这些颗粒因标记特别强烈而突出。这些颗粒最常出现在肌膜下区域的观察结果表明,它们可能是正在进行胞吐作用的成熟颗粒。因此,在内分泌心肌细胞分泌颗粒上发现的G(o)α可能在心脏激素的调节性胞吐作用中发挥作用。G(o)α在心房和心室肌细胞中的肌膜定位支持了G(o)α在跨膜信号转导中的作用。此外,G(o)α的小窝定位可能为整合G(o)介导的信号事件提供一个分隔基础。