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培养的大鼠肌管上乙酰胆碱受体的流动性和去污剂可提取性:一种相关性

Mobility and detergent extractability of acetylcholine receptors on cultured rat myotubes: a correlation.

作者信息

Stya M, Axelrod D

出版信息

J Cell Biol. 1983 Jul;97(1):48-51. doi: 10.1083/jcb.97.1.48.

Abstract

On aneurally cultured rat primary myotubes, 10% of the acetylcholine receptors (AChR) are found aggregated and immobilized in endogenous clusters. The remaining receptors are diffusely distributed over the cell membrane and the majority of these are free to diffuse in the plane of the membrane. This study correlates the mobility of AChR (as measured with the fluorescence photobleaching recovery technique, FPR) with the detergent extractability of this receptor. Gentle detergent extraction of the cells removes the lipid membrane and the soluble cytoplasmic proteins but leaves an intact cytoskeletal framework on the substrate. Two studies indicate a correlation between mobility and extractability: (a) mobility of diffusely distributed AChR decreases as myotubes age in culture; previous work showed that extractability of AChR decreases as myotubes age in culture (Prives, J., C. Christian, S. Penman, and K. Olden, 1980, In Tissue Culture in Neurobiology, E. Giacobini, A. Vernadakis, and A. Shahar, editors, Raven Press, New York, 35-52); (b) mobility of clustered AChR increases when cells are treated with metabolic inhibitors such as sodium azide (NaN3); extractability of clustered AChR also increases with this treatment. From these results we suggest the involvement of a cytoskeletal framework in the immobilization of AChR on the cell surface.

摘要

在无神经培养的大鼠原代肌管上,发现10%的乙酰胆碱受体(AChR)聚集并固定在内源性簇中。其余的受体则分散分布在细胞膜上,其中大多数可以在膜平面内自由扩散。本研究将AChR的流动性(用荧光光漂白恢复技术,即FPR测量)与该受体的去污剂可提取性联系起来。用温和的去污剂提取细胞可去除脂质膜和可溶性细胞质蛋白,但在底物上留下完整的细胞骨架框架。两项研究表明了流动性和可提取性之间的相关性:(a)随着培养的肌管老化,分散分布的AChR的流动性降低;先前的研究表明,随着培养的肌管老化,AChR的可提取性降低(普里夫斯,J.,C.克里斯蒂安,S.彭曼,和K.奥尔登,1980年,载于《神经生物学中的组织培养》,E.贾科比尼、A.韦尔纳达基斯和A.沙哈尔编,拉文出版社,纽约,第35 - 52页);(b)当用代谢抑制剂如叠氮化钠(NaN₃)处理细胞时,成簇AChR的流动性增加;这种处理也会增加成簇AChR的可提取性。根据这些结果,我们认为细胞骨架框架参与了AChR在细胞表面的固定。

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