Choi J, Costa M L, Mermelstein C S, Chagas C, Holtzer S, Holtzer H
Department of Biochemistry, University of Pennsylvania, Philadelphia 19104.
Proc Natl Acad Sci U S A. 1990 Oct;87(20):7988-92. doi: 10.1073/pnas.87.20.7988.
Shortly after their birth, postmitotic mononucleated myoblasts in myotomes, limb buds, and conventional muscle cultures elongate and assemble a cohort of myofibrillar proteins into definitively striated myofibrils. MyoD induces a number of immortalized and/or transformed nonmuscle cells to express desmin and several myofibrillar proteins and to fuse into myosacs. We now report that MyoD converts normal dermal fibroblasts, chondroblasts, gizzard smooth muscle, and pigmented retinal epithelial cells into elongated postmitotic mononucleated striated myoblasts. The sarcomeric localization of antibodies to desmin, alpha-actinin, titin, troponin-I, alpha-actin, myosin heavy chain, and myomesin in these converted myoblasts are indistinguishable from in vivo and in vitro normal myoblasts. Converted myoblasts fuse into typical anisodiametric multinucleated myotubes that often contract spontaneously. Conversion and subsequent expression of the skeletal myogenic program are autonomous events, occurring in four nonmuscle microenvironments consisting of different combinations of foreign extracellular matrix molecules. Early events associated with conversion by MyoD involve (i) withdrawal from the cell cycle, (ii) down-regulation of the subverted cell's ongoing differentiation program, and (iii) initiation of desmin synthesis in presumptive myoblasts and dramatic redistribution of microtubules and desmin intermediate filaments in postmitotic myoblasts.
在出生后不久,体节、肢芽和传统肌肉培养物中的有丝分裂后单核成肌细胞会伸长,并将一群肌原纤维蛋白组装成明确横纹的肌原纤维。MyoD诱导一些永生化和/或转化的非肌肉细胞表达结蛋白和几种肌原纤维蛋白,并融合形成肌囊。我们现在报告,MyoD可将正常的真皮成纤维细胞、软骨细胞、砂囊平滑肌细胞和色素性视网膜上皮细胞转化为伸长的有丝分裂后单核横纹成肌细胞。在这些转化的成肌细胞中,针对结蛋白、α-辅肌动蛋白、肌联蛋白、肌钙蛋白-I、α-肌动蛋白、肌球蛋白重链和肌间线蛋白的抗体的肌节定位与体内和体外正常成肌细胞中的定位没有区别。转化的成肌细胞融合形成典型的不等径多核肌管,这些肌管常常会自发收缩。骨骼肌生成程序的转化及随后的表达是自主事件,发生在由不同组合的外源细胞外基质分子组成的四种非肌肉微环境中。与MyoD转化相关的早期事件包括:(i)退出细胞周期;(ii)下调被颠覆细胞正在进行的分化程序;(iii)在假定的成肌细胞中启动结蛋白合成,并在有丝分裂后成肌细胞中使微管和结蛋白中间丝发生显著重新分布。