Barrasso Anthony P, Tong Xuefei, Poché Ross A
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, Texas 77030.
Program in Integrative Molecular and Biomedical Sciences, Baylor College of Medicine, Houston, Texas 77030.
Genesis. 2018 Feb;56(2). doi: 10.1002/dvg.23087. Epub 2017 Dec 27.
Mitochondria are incredibly dynamic organelles that undergo continuous fission and fusion events to control morphology, which profoundly impacts cell physiology including cell cycle progression. This is highlighted by the fact that most major human neurodegenerative diseases are due to specific disruptions in mitochondrial fission or fusion machinery and null alleles of these genes result in embryonic lethality. To gain a better understanding of the pathophysiology of such disorders, tools for the in vivo assessment of mitochondrial dynamics are required. It would be particularly advantageous to simultaneously image mitochondrial fission-fusion coincident with cell cycle progression. To that end, we have generated a new transgenic reporter mouse, called mito::mKate2 that ubiquitously expresses a mitochondria localized far-red mKate2 fluorescent protein. Here we show that mito::mKate2 mice are viable and fertile and that mKate2 fluorescence can be spectrally separated from the previously developed Fucci cell cycle reporters. By crossing mito::mKate2 mice to the ROSA26R-mTmG dual fluorescent Cre reporter line, we also demonstrate the potential utility of mito::mKate2 for genetic mosaic analysis of mitochondrial phenotypes.
线粒体是极其动态的细胞器,会经历持续的分裂和融合事件以控制形态,这对包括细胞周期进程在内的细胞生理有着深远影响。这一点体现在大多数主要的人类神经退行性疾病是由于线粒体分裂或融合机制的特定破坏,而这些基因的无效等位基因会导致胚胎致死。为了更好地理解此类疾病的病理生理学,需要用于体内评估线粒体动态的工具。同时对与细胞周期进程同时发生的线粒体分裂 - 融合进行成像将特别有利。为此,我们培育了一种新的转基因报告小鼠,称为mito::mKate2,它普遍表达一种定位于线粒体的远红色mKate2荧光蛋白。在这里我们表明,mito::mKate2小鼠是有活力且可育的,并且mKate2荧光可以与先前开发的Fucci细胞周期报告基因在光谱上分离。通过将mito::mKate2小鼠与ROSA26R - mTmG双荧光Cre报告基因系杂交,我们还证明了mito::mKate2在线粒体表型的基因镶嵌分析中的潜在用途。