首页> 外文期刊>Cell biology international. >Permeabilization by streptolysin-o reveals a role for calcium-dependent protein kinase c isoforms alpha and beta in the response of cultured cardiomyocytes to hyposmotic challenge.
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Permeabilization by streptolysin-o reveals a role for calcium-dependent protein kinase c isoforms alpha and beta in the response of cultured cardiomyocytes to hyposmotic challenge.

机译:链球菌溶血素-o的透化作用揭示了钙依赖性蛋白激酶c同工型alpha和beta在培养的心肌细胞对低渗挑战的反应中的作用。

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摘要

Immunocytochemical techniques indicate that the uninhibited activity of protein kinase C alpha and protein kinase C beta are necessary for a normal regulatory volume decrease (RVD) response of cultured chick embryo cardiomyocytes subjected to a hyposmotic environment. Antibodies against protein kinase C isoforms alpha, beta, gamma and epsilon were introduced into the cultured myocytes using a developed streptolysin-O (SLO) permeabilization technique that allows the targeted cells to accumulate large biomolecules without perturbing their normal physiological state. The loaded cells were then tested for their ability to RVD when submitted to hypo-osmotic stimulus. Results show that exposing the cultured cells to SLO in the presence of antibodies against protein kinase C alpha and beta, prior to volume challenge, significantly slows the RVD rate. Additional experiments that combined anti-alpha and anti-beta antibodies in the same exposure media did not result in a significantly different rate than the anti-alpha or anti-beta rates alone. The evidence gained in this study is in agreement with previous work in the cultured chick embryo cardiomyocyte that report the involvement of a calcium dependent protein kinase C in the signal transduction pathway of the RVD. Copyright 1999 Academic Press.
机译:免疫细胞化学技术表明,蛋白激酶Cα和蛋白激酶Cβ的抑制活性对于在低渗环境下培养的鸡胚心肌细胞的正常调节体积减少(RVD)反应是必需的。使用发达的链球菌溶血素-O(SLO)透化技术,将针对蛋白激酶C亚型的抗体α,β,γ和ε引入培养的心肌细胞,该技术可使靶细胞积聚大分子而不干扰其正常生理状态。然后,当受到低渗透刺激时,测试加载的细胞的RVD能力。结果表明,在体积挑战之前,在存在针对蛋白激酶Cα和β的抗体的情况下,将培养的细胞暴露于SLO会显着降低RVD速率。在相同的暴露介质中结合使用抗α和抗β抗体的其他实验所产生的比率与单独使用抗α或抗β的比率没有明显不同。在这项研究中获得的证据与先前在培养的鸡胚心肌细胞中的研究相一致,该研究报道了钙依赖性蛋白激酶C参与RVD的信号转导途径。版权所有1999,学术出版社。

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