Dlugosz A A, Antin P B, Nachmias V T, Holtzer H
J Cell Biol. 1984 Dec;99(6):2268-78. doi: 10.1083/jcb.99.6.2268.
The topographical relationship between stress fiber-like structures (SFLS) and nascent myofibrils was examined in cultured chick cardiac myocytes by immunofluorescence microscopy. Antibodies against muscle-specific light meromyosin (anti-LMM) and desmin were used to distinguish cardiac myocytes from fibroblastic cells. By various combinations of staining with rhodamine-labeled phalloidin, anti-LMM, and antibodies against chick brain myosin and smooth muscle alpha-actinin, we observed the following relationships between transitory SFLS and nascent and mature myofibrils: (a) more SFLS were present in immature than mature myocytes; (b) in immature myocytes a single fluorescent fiber would stain as a SFLS distally and as a striated myofibril proximally, towards the center of the cell; (c) in regions of a myocyte not yet penetrated by the elongating myofibrils, SFLS were abundant; and (d) in regions of a myocyte with numerous mature myofibrils, SFLS had totally disappeared. Spontaneously contracting striated myofibrils with definitive Z-band regions were present long before anti-desmin localized in the I-Z-band region and long before morphologically recognizable structures periodically link Z-bands to the sarcolemma. These results suggest a transient one-on-one relationship between individual SFLS and newly emerging individual nascent myofibrils. Based on these and other relevant data, a complex, multistage molecular model is presented for myofibrillar assembly and maturation. Lastly, it is of considerable theoretical interest to note that mature cardiac myocytes, like mature skeletal myotubes, lack readily detectable stress fibers.
通过免疫荧光显微镜检查,研究了培养的鸡心肌细胞中应力纤维样结构(SFLS)与新生肌原纤维之间的拓扑关系。使用针对肌肉特异性轻酶解肌球蛋白(抗LMM)和结蛋白的抗体来区分心肌细胞和成纤维细胞。通过用罗丹明标记的鬼笔环肽、抗LMM以及针对鸡脑肌球蛋白和平滑肌α-肌动蛋白的抗体进行各种染色组合,我们观察到了短暂性SFLS与新生及成熟肌原纤维之间的以下关系:(a)未成熟心肌细胞中的SFLS比成熟心肌细胞中的更多;(b)在未成熟心肌细胞中,单个荧光纤维在远端会被染成SFLS,而在近端朝着细胞中心会被染成横纹肌原纤维;(c)在肌原纤维尚未延伸到的心肌细胞区域,SFLS丰富;(d)在有许多成熟肌原纤维的心肌细胞区域,SFLS完全消失。具有明确Z带区域的自发收缩横纹肌原纤维早在抗结蛋白定位于I-Z带区域之前以及早在形态学上可识别的结构将Z带与肌膜周期性连接之前就已存在。这些结果表明单个SFLS与新出现的单个新生肌原纤维之间存在短暂的一对一关系。基于这些及其他相关数据,提出了一个关于肌原纤维组装和成熟的复杂多阶段分子模型。最后,值得注意的是,成熟心肌细胞与成熟骨骼肌管一样,缺乏易于检测到的应力纤维,这在理论上具有相当大的意义。