Dynamics of myogenic differentiation using a novel Myogenin knock-in reporter mouse

IF 5.3 2区医学 Q2 CELL BIOLOGY

Skeletal Muscle Pub Date : 2020-12-21 DOI:10.1101/2020.12.21.423736

Maria Benavente-Diaz, Glenda Comai, D. Di Girolamo, Francina Langa, S. Tajbakhsh

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引用次数: 7

Abstract

Background Myogenin is a transcription factor that is expressed during terminal myoblast differentiation in embryonic development and adult muscle regeneration. Investigation of this cell state transition has been hampered by the lack of a sensitive reporter to dynamically track cells during differentiation. Results Here, we report a knock-in mouse line expressing the tdTOMATO fluorescent protein from the endogenous Myogenin locus. Expression of tdTOMATO in Myog ntdTom mice recapitulated endogenous Myogenin expression during embryonic muscle formation and adult regeneration and enabled the isolation of the MYOGENIN + cell population. We also show that tdTOMATO fluorescence allows tracking of differentiating myoblasts in vitro and by intravital imaging in vivo. Lastly, we monitored by live imaging the cell division dynamics of differentiating myoblasts in vitro and showed that a fraction of the MYOGENIN + population can undergo one round of cell division, albeit at a much lower frequency than MYOGENIN − myoblasts. Conclusions We expect that this reporter mouse will be a valuable resource for researchers investigating skeletal muscle biology in developmental and adult contexts.

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使用新型肌原蛋白敲入报告小鼠的肌源性分化动力学

背景肌生成素是一种在胚胎发育和成年肌肉再生的成肌细胞分化末期表达的转录因子。由于缺乏在分化过程中动态跟踪细胞的敏感报告子，对这种细胞状态转变的研究受到阻碍。结果在这里，我们报道了一个从内源性肌生成素基因座表达tdTOMATO荧光蛋白的敲除小鼠系。tdTOMATO在Myog ntdTom小鼠中的表达再现了胚胎肌肉形成和成年再生过程中内源性肌生成素的表达，并能够分离MYGENIN+细胞群。我们还表明，tdTOMATO荧光可以在体外和体内通过活体内成像跟踪分化的成肌细胞。最后，我们通过实时成像监测了体外分化成肌细胞的细胞分裂动力学，并表明一部分MYGENIN+群体可以经历一轮细胞分裂，尽管频率远低于MYGENIN-成肌细胞。结论我们期望这种报告小鼠将成为研究人员在发育和成年背景下研究骨骼肌生物学的宝贵资源。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Skeletal Muscle CELL BIOLOGY-

CiteScore

9.10

自引率

0.00%

发文量

审稿时长

12 weeks

期刊介绍： The only open access journal in its field, Skeletal Muscle publishes novel, cutting-edge research and technological advancements that investigate the molecular mechanisms underlying the biology of skeletal muscle. Reflecting the breadth of research in this area, the journal welcomes manuscripts about the development, metabolism, the regulation of mass and function, aging, degeneration, dystrophy and regeneration of skeletal muscle, with an emphasis on understanding adult skeletal muscle, its maintenance, and its interactions with non-muscle cell types and regulatory modulators. Main areas of interest include: -differentiation of skeletal muscle- atrophy and hypertrophy of skeletal muscle- aging of skeletal muscle- regeneration and degeneration of skeletal muscle- biology of satellite and satellite-like cells- dystrophic degeneration of skeletal muscle- energy and glucose homeostasis in skeletal muscle- non-dystrophic genetic diseases of skeletal muscle, such as Spinal Muscular Atrophy and myopathies- maintenance of neuromuscular junctions- roles of ryanodine receptors and calcium signaling in skeletal muscle- roles of nuclear receptors in skeletal muscle- roles of GPCRs and GPCR signaling in skeletal muscle- other relevant aspects of skeletal muscle biology. In addition, articles on translational clinical studies that address molecular and cellular mechanisms of skeletal muscle will be published. Case reports are also encouraged for submission. Skeletal Muscle reflects the breadth of research on skeletal muscle and bridges gaps between diverse areas of science for example cardiac cell biology and neurobiology, which share common features with respect to cell differentiation, excitatory membranes, cell-cell communication, and maintenance. Suitable articles are model and mechanism-driven, and apply statistical principles where appropriate; purely descriptive studies are of lesser interest.

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