斑马鱼胚胎中快肌细胞的融合需要成肌素。
Myomaker is required for the fusion of fast-twitch myocytes in the zebrafish embryo.
作者信息
Zhang Weibin, Roy Sudipto
机构信息
Institute of Molecular and Cell Biology, Proteos, 61 Biopolis Drive, Singapore 138673, Singapore.
Institute of Molecular and Cell Biology, Proteos, 61 Biopolis Drive, Singapore 138673, Singapore; Department of Pediatrics, Yong Loo Lin School of Medicine, National University of Singapore, 1E Kent Ridge Road, Singapore 119288, Singapore; Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore.
出版信息
Dev Biol. 2017 Mar 1;423(1):24-33. doi: 10.1016/j.ydbio.2017.01.019. Epub 2017 Feb 1.
During skeletal muscle development, myocytes aggregate and fuse to form multinucleated muscle fibers. Inhibition of myocyte fusion is thought to significantly derail the differentiation of functional muscle fibers. Despite the purported importance of fusion in myogenesis, in vivo studies of this process in vertebrates are rather limited. Myomaker, a multipass transmembrane protein, has been shown to be the first muscle-specific fusion protein essential for myocyte fusion in the mouse. We have generated loss-of-function alleles in zebrafish myomaker, and found that fusion of myocytes into syncytial fast-twitch muscles was significantly compromised. However, mutant myocytes could be recruited to fuse with wild-type myocytes in chimeric embryos, albeit rather inefficiently. Conversely, overexpression of Myomaker was sufficient to induce hyperfusion among fast-twitch myocytes, and it also induced fusion among slow-twitch myocytes that are normally fusion-incompetent. In line with this, Myomaker overexpression also triggered fusion in another myocyte fusion mutant compromised in the function of the junctional cell adhesion molecule, Jam2a. We also provide evidence that Rac, a regulator of actin cytoskeleton, requires Myomaker activity to induce fusion, and that an approximately 3kb of myomaker promoter sequence, with multiple E-box motifs, is sufficient to direct expression within the fast-twitch muscle lineage. Taken together, our findings underscore a conserved role for Myomaker in vertebrate myocyte fusion. Strikingly, and in contrast to the mouse, homozygous myomaker mutants are viable and do not exhibit discernible locomotory defects. Thus, in the zebrafish, myocyte fusion is not an absolute requirement for skeletal muscle morphogenesis and function.
在骨骼肌发育过程中,肌细胞聚集并融合形成多核肌纤维。肌细胞融合的抑制被认为会严重阻碍功能性肌纤维的分化。尽管融合在肌生成中据称很重要,但脊椎动物体内对这一过程的研究相当有限。肌生成素是一种多次跨膜蛋白,已被证明是小鼠肌细胞融合所必需的首个肌肉特异性融合蛋白。我们在斑马鱼中产生了肌生成素功能缺失等位基因,发现肌细胞融合成多核快速收缩肌的过程受到显著影响。然而,在嵌合胚胎中,突变的肌细胞可以被招募与野生型肌细胞融合,尽管效率相当低。相反,肌生成素的过表达足以诱导快速收缩肌细胞之间的过度融合,并且还能诱导通常无融合能力的慢速收缩肌细胞之间的融合。与此一致的是,肌生成素的过表达也在另一个因连接细胞粘附分子Jam2a功能受损的肌细胞融合突变体中引发了融合。我们还提供证据表明,肌动蛋白细胞骨架的调节因子Rac需要肌生成素的活性来诱导融合,并且一个约3kb带有多个E-box基序的肌生成素启动子序列足以指导在快速收缩肌谱系中的表达。综上所述,我们的研究结果强调了肌生成素在脊椎动物肌细胞融合中的保守作用。引人注目的是,与小鼠不同,纯合肌生成素突变体是存活的,并且没有表现出明显的运动缺陷。因此,在斑马鱼中,肌细胞融合并非骨骼肌形态发生和功能的绝对必要条件。