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Am J Physiol Cell Physiol (July 2, 2003). doi:10.1152/ajpcell.00104.2003
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Submitted on March 19, 2003
Accepted on June 26, 2003

MUSCLE ELECTROTRANSFER AS A TOOL FOR STUDYING MUSCLE FIBER-SPECIFICITY AND NERVE-DEPENDENT ACTIVITY OF PROMOTERS

Anne BERTRAND1, Valerie NGO-MULLER1, Daniele HENTZEN1, Jean-Paul CONCORDET1, Dominique DAEGELEN1*, and David TUIL1

1 Departement Genetique, Developpement et Pathologie Moleculaire, Institut Cochin, PARIS, France

* To whom correspondence should be addressed. E-mail: daegelen{at}cochin.inserm.fr.

Muscle electrotransfer has recently become a promising tool for efficient delivery of plasmids and transgene expression in skeletal muscle. So far, this technology has been mainly applied in order to use muscle as a bioreactor for production of therapeutic proteins. However, it remains to be determined whether muscle electrotransfer may also be accurately used as an alternative tool to transgenesis for studying some aspects of muscle-specific gene control that need to be explored in the in vivo context of fully mature muscle fibers, such as fiber-specificity and nerve-dependency. It was also not known to what extent the initial electrical stimulations may alter muscle physiology and gene expression. Therefore, optimized conditions of skeletal muscle electroporation were first tested for their effects on muscles of transgenic mice harboring a pM310-CAT transgene in which the CAT reporter gene was under control of the fast IIB fiber-specific and nerve-dependent aldolase A pM promoter. Surprisingly, electrostimulation led to a drastic but transient shutdown of pM310-CAT transgene expression that was concomitant with a very transient activation of MyoD and mostly, with activation of myogenin, suggesting profound alterations in the transcriptional status of the electroporated muscle. Return to a normal transcriptional state was observed 7 to 10 days after electroporation. Therefore, we investigated whether a reporter construct placed under control of pM could exhibit fiber-specific expression when analyzed 10 days after electrotransfer in either fast tibialis anterior or slow soleus. Here we show that not only fiber specificity, but also nerve dependency of a pM driven construct could be reproduced. However, following electrotransfer, pM displayed a less tight control than what was previously observed for the same promoter when integrated in a chromatin context.




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