构建一个MyoD基因敲入报告基因小鼠品系以研究肌肉干细胞动力学和异质性。
Generation of a MyoD knock-in reporter mouse line to study muscle stem cell dynamics and heterogeneity.
作者信息
Fujita Ryo, Mizuno Seiya, Sadahiro Taketaro, Hayashi Takuto, Sugasawa Takehito, Sugiyama Fumihiro, Ono Yusuke, Takahashi Satoru, Ieda Masaki
机构信息
Division of Regenerative Medicine, Transborder Medical Research Center, Institute of Medicine, University of Tsukuba, Ibaraki 305-8575, Japan.
Department of Cardiology, Institute of Medicine, University of Tsukuba, Ibaraki 305-8575, Japan.
出版信息
iScience. 2023 Apr 8;26(5):106592. doi: 10.1016/j.isci.2023.106592. eCollection 2023 May 19.
Myoblast determination protein 1 (MyoD) dynamics define the activation status of muscle stem cells (MuSCs), aiding in muscle tissue regeneration after injury. However, the lack of experimental platforms to monitor MyoD dynamics and has hampered the investigation of fate determination and heterogeneity of MuSCs. Herein, we report a MyoD knock-in (MyoD-KI) reporter mouse expressing tdTomato at the endogenous locus. Expression of tdTomato in MyoD-KI mice recapitulated the endogenous MyoD expression dynamics and during the early phase of regeneration . Additionally, we showed that tdTomato fluorescence intensity defines MuSC activation status without immunostaining. Based on these features, we developed a high-throughput screening system to assess the effects of drugs on the behavior of MuSCs . Thus, MyoD-KI mice are an invaluable resource for studying the dynamics of MuSCs, including their fate decisions and heterogeneity, and for drug screening in stem cell therapy.
成肌细胞决定蛋白1（MyoD）的动态变化定义了肌肉干细胞（MuSCs）的激活状态，有助于损伤后肌肉组织的再生。然而，缺乏监测MyoD动态变化的实验平台阻碍了对MuSCs命运决定和异质性的研究。在此，我们报告了一种在MyoD基因座敲入（MyoD-KI）报告基因的小鼠，其在内源位点表达tdTomato。tdTomato在MyoD-KI小鼠中的表达重现了内源性MyoD的表达动态，且在再生早期亦是如此。此外，我们表明tdTomato荧光强度无需免疫染色即可定义MuSC的激活状态。基于这些特性，我们开发了一种高通量筛选系统来评估药物对MuSCs行为的影响。因此，MyoD-KI小鼠是研究MuSCs动态变化（包括其命运决定和异质性）以及进行干细胞治疗药物筛选的宝贵资源。
Zotero 插件