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首页> 外文期刊>American Journal of Physiology >Real-time imaging of peroxisome proliferator-activated receptor-gamma coactivator-1alpha promoter activity in skeletal muscles of living mice.
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Real-time imaging of peroxisome proliferator-activated receptor-gamma coactivator-1alpha promoter activity in skeletal muscles of living mice.

机译：活体小鼠骨骼肌中过氧化物酶体增殖物激活的受体-γcoactivator-1alpha启动子活性的实时成像。

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摘要

In response to sustained increase in contractile activity, mammalian skeletal muscle undergoes adaptation with enhanced mitochondrial biogenesis and fiber type switching. The peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha) was recently identified as a key regulator for these adaptive processes. To investigate the sequence elements in the PGC-1alpha gene that are responsible for activity-dependent transcriptional activation, we have established a unique system to analyze promoter activity in skeletal muscle of living mice. Expression of PGC-1alpha-firefly luciferase reporter gene in mouse tibialis anterior muscle transfected by electric pulse-mediated gene transfer was assessed repeatedly in the same muscle by using optical bioluminescence imaging analysis before and after low-frequency (10 Hz) motor nerve stimulation. Nerve stimulation (2 h) resulted in a transient 3-fold increase (P < 0.05) in PGC-1alpha promoter activity along with a 1.6-fold increase (P < 0.05) in endogenous PGC-1alpha mRNA. Mutation of two consensus myocyte enhancer factor 2 (MEF2) binding sites (-2901 and -1539) or a cAMP response element (CRE) (-222) completely abolished nerve stimulation-induced increase in PGC-1alpha promoter activity. These findings provide direct evidence that contractile activity-induced PGC-1alpha promoter activity in skeletal muscle is dependent on the MEF2 and the CRE sequence elements. The experimental methods used in the present study have general applicability to studies of gene regulation in muscle.
机译：为了响应持续增加的收缩活动，哺乳动物骨骼肌经历了线粒体生物发生增强和纤维类型转换的适应。过氧化物酶体增殖物激活受体-γ共激活因子1alpha（PGC-1alpha）最近被确定为这些适应性过程的关键调节器。为了研究PGC-1alpha基因中负责活性依赖性转录激活的序列元件，我们建立了一个独特的系统来分析活体小鼠骨骼肌中启动子的活性。在低频（10 Hz）运动神经刺激前后，通过光学生物发光成像分析，通过光学生物发光成像分析，反复评估了电脉冲介导的基因转移转染的小鼠胫前肌中PGC-1alpha-萤火虫荧光素酶报告基因的表达。神经刺激（2小时）导致PGC-1alpha启动子活性瞬时增加3倍（P <0.05），而内源性PGC-1alpha mRNA增加1.6倍（P <0.05）。两个共有肌细胞增强因子2（MEF2）结合位点（-2901和-1539）或cAMP反应元件（CRE）（-222）的突变完全消除了神经刺激引起的PGC-1alpha启动子活性的增加。这些发现提供了直接的证据，即骨骼肌中的收缩活性诱导的PGC-1alpha启动子活性取决于MEF2和CRE序列元件。本研究中使用的实验方法普遍适用于肌肉中基因调控的研究。

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