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Am J Physiol Cell Physiol (May 19, 2004). doi:10.1152/ajpcell.00425.2003
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Submitted on October 2, 2003
Accepted on May 13, 2004

Real-time Imaging of Peroxisome Proliferator Activated Receptor {gamma} Co-activator-1{alpha} Promoter Activity in Skeletal Muscles of Living Mice

Takayuki Akimoto1, Brian S Sorg2, and Zhen Yan1*

1 Medicine, Duke University Medical Center, Durham, NC, USA
2 Radiation Oncology, Duke University Medical Center, Durham, NC, USA

* To whom correspondence should be addressed. E-mail: zhen.yan{at}duke.edu.

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} co-activator-1{alpha} (PGC-1{alpha}) has recently been identified as a key regulator for these adaptive processes. To investigate the sequence elements in the PGC-1{alpha} 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-1{alpha}-firefly luciferase reporter gene in mouse tibialis anterior (TA) muscle transfected by electric pulse-mediated gene transfer was assessed repeatedly in the same muscle by 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-1{alpha} promoter activity along with a 1.6-fold increase (P<0.05) in endogenous PGC-1{alpha} 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-1{alpha} promoter activity. These findings provide direct evidence that contractile activity-induced PGC-1{alpha} promoter activity in skeletal muscle is dependent on the MEF2 and the CRE sequence elements. The experimental methods used here have general applicability to studies of gene regulation in muscle.




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