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通过与HVJ(仙台病毒)融合形成的骨骼肌成肌细胞与体细胞之间异核体的特性;对肌源性分化的影响。

Characterization of heterokaryons between skeletal myoblasts and somatic cells formed by fusion with HVJ (Sendai virus); effects on myogenic differentiation.

作者信息

Hirayama E, Udaka Y, Kawai T, Kim J

机构信息

Institutes of Molecular and Cellular Biology for Pharmaceutical Sciences, Kyoto Pharmaceutical University, Japan.

出版信息

Cell Struct Funct. 2001 Feb;26(1):37-47. doi: 10.1247/csf.26.37.

Abstract

In skeletal myogenic differentiation, myoblasts fuse with myogenic cells spontaneously, but do not fuse with non-myogenic cells either in vivo or in vitro, suggesting that the fusion of myoblasts with non-myogenic cells is unsuitable for differentiation. To understand the inevitability of the fusion among myoblasts, we prepared heterokaryons in crosses between quail myoblasts transformed with a temperature-sensitive mutant of Rous sarcoma virus (QM-RSV cells) and rodent non-myogenic cells, such as tumor cells, fibroblasts, or neurogenic cells by HVJ (Sendai virus) and examined how myogenic differentiation was influenced in the prepared heterokaryons, focusing on myogenin expression and myofibril formation as markers of differentiation. When presumptive QM-RSV cells were fused with non-myogenic cells by HVJ and induced to differentiate, both myogenin expression and myofibril formation were suppressed. When myotubes of QM-RSV cells that had already expressed myogenin and formed myofibrils were fused with non-myogenic cells, both myogenin and myofibrils disappeared. Especially, fibrous structures of myofibrils were significantly lost and dots or aggregations of F-actin were formed within 24 hr after formation of heterokaryons. However, the fusion of presumptive or differentiated QM-RSV cells with rodent myoblasts did not disturb myogenin expression or myofibril formation. These results suggest that mutual fusion of myoblasts is indispensable for normal myogenic differentiation irrespective of the species, and that some factors inhibiting myogenic differentiation exist in the cytoplasm of non-myogenic cells, but not in myoblasts.

摘要

在骨骼肌生肌分化过程中,成肌细胞会自发地与肌源性细胞融合,但在体内或体外均不会与非肌源性细胞融合,这表明成肌细胞与非肌源性细胞的融合不适合分化。为了理解成肌细胞之间融合的必然性,我们通过HVJ(仙台病毒)在经劳斯肉瘤病毒温度敏感突变体转化的鹌鹑成肌细胞(QM-RSV细胞)与啮齿动物非肌源性细胞(如肿瘤细胞、成纤维细胞或神经源性细胞)之间的杂交中制备了异核体,并研究了在制备的异核体中肌源性分化是如何受到影响的,重点关注作为分化标志物的肌细胞生成素表达和肌原纤维形成。当假定的QM-RSV细胞通过HVJ与非肌源性细胞融合并诱导分化时,肌细胞生成素表达和肌原纤维形成均受到抑制。当已经表达肌细胞生成素并形成肌原纤维的QM-RSV细胞的肌管与非肌源性细胞融合时,肌细胞生成素和肌原纤维均消失。特别是,在异核体形成后24小时内,肌原纤维的纤维结构明显丧失,F-肌动蛋白形成点状或聚集物。然而,假定的或分化的QM-RSV细胞与啮齿动物成肌细胞的融合并未干扰肌细胞生成素表达或肌原纤维形成。这些结果表明,无论物种如何,成肌细胞的相互融合对于正常的肌源性分化都是必不可少的,并且在非肌源性细胞的细胞质中存在一些抑制肌源性分化的因子,而成肌细胞中则不存在。

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