McKenna N, Meigs J B, Wang Y L
J Cell Biol. 1985 Jan;100(1):292-6. doi: 10.1083/jcb.100.1.292.
We have investigated whether living muscle and nonmuscle cells can discriminate between microinjected muscle and nonmuscle actins. Muscle actin purified from rabbit back and leg muscles and labeled with fluorescein isothiocyanate, and nonmuscle actin purified from lamb brain and labeled with lissamine rhodamine B sulfonyl chloride, were co-injected into chick embryonic cardiac myocytes and fibroblasts. When fluorescence images of the two actins were compared using filter sets selective for either fluorescein isothiocyanate or lissamine rhodamine B sulfonyl chloride, essentially identical patterns of distribution were detected in both muscle and nonmuscle cells. In particular, we found no structure that, at this level of resolution, shows preferential binding of muscle or nonmuscle actin. In fibroblasts, both actins are associated primarily with stress fibers and ruffles. In myocytes, both actins are localized in sarcomeres. In addition, the distribution of structures containing microinjected actins is similar to that of structure containing endogenous F-actin, as revealed by staining with fluorescent phalloidin or phallacidin. Our results suggest that, at least under these experimental conditions, actin-binding sites in muscle and nonmuscle cells do not discriminate among different forms of actins.
我们研究了活的肌肉细胞和非肌肉细胞是否能够区分显微注射的肌肉肌动蛋白和非肌肉肌动蛋白。从兔背部和腿部肌肉中纯化并标记有异硫氰酸荧光素的肌肉肌动蛋白,以及从羊脑中纯化并标记有丽丝胺罗丹明B磺酰氯的非肌肉肌动蛋白,被共同注射到鸡胚心肌细胞和成纤维细胞中。当使用对异硫氰酸荧光素或丽丝胺罗丹明B磺酰氯有选择性的滤光片组比较两种肌动蛋白的荧光图像时,在肌肉细胞和非肌肉细胞中都检测到了基本相同的分布模式。特别是,在这个分辨率水平上,我们没有发现显示肌肉或非肌肉肌动蛋白优先结合的结构。在成纤维细胞中,两种肌动蛋白主要与应力纤维和褶皱相关。在心肌细胞中,两种肌动蛋白都定位于肌节中。此外,如用荧光鬼笔环肽或鬼笔环肽染色所显示的,含有显微注射肌动蛋白的结构的分布与含有内源性F-肌动蛋白的结构的分布相似。我们的结果表明,至少在这些实验条件下,肌肉细胞和非肌肉细胞中的肌动蛋白结合位点不会区分不同形式的肌动蛋白。