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 : 2021-02-18 DOI:10.1186/s13395-021-00260-x

Maria Benavente-Diaz, Glenda Comai, Daniela Di Girolamo, Francina Langa, Shahragim Tajbakhsh

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引用次数: 3

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:肌原素是一种转录因子，在胚胎发育和成人肌肉再生的成肌细胞末梢分化过程中表达。由于缺乏敏感的报告细胞动态跟踪分化过程，对这种细胞状态转变的研究一直受到阻碍。结果:在这里，我们报道了一个从内源性肌原蛋白位点表达tdTOMATO荧光蛋白的敲入小鼠系。tdTOMATO在MyogntdTom小鼠中的表达重现了胚胎肌肉形成和成体再生过程中内源性Myogenin的表达，并使Myogenin +细胞群的分离成为可能。我们还表明，tdTOMATO荧光可以在体外和体内活体成像中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，结果表明，一小部分MYOGENIN+群体可以经历一轮细胞分裂，尽管频率远低于MYOGENIN-成肌细胞。结论:我们期望这只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.