Analysis of neural-responsive myogenin upstream sequences by myoblast implantation

Developmental Biology
G Gibney, A Buonanno

Abstract

The expression of myogenin is suppressed during innervation and has been implicated in determining properties of skeletal muscle which are regulated by electrical activity. We previously reported that transcription driven by 3700 bp of the mouse myogenin upstream sequence (MYG3700) is activated by denervation in transgenic mice (Nucleic Acids Res. 21, 5684-5693, 1993). To extend our investigation of the activity dependence of the myogenin promoter, we have utilized myoblast implantation as a novel approach to in vivo reporter analysis. Myoblasts for hindlimb injections were generated by stable transfection of chloramphenicol acetyltransferase (CAT) reporters into a beta-galactosidase-expressing line of C2 cells. In vitro characterization of stable myoblast clones carrying myogenin-CAT deletion constructs revealed that while the proximal myogenin 5'-flanking sequence confers myotube specificity, high-level expression requires a region upstream (-335 to -1102) which depends on chromosomal integration for its function. For analysis by implantation, incorporation of injected myoblasts into existing myofibers was confirmed by histochemical staining. Using clonal myoblasts harboring nicotinic receptor alpha-subunit (alpha 800) and my...Continue Reading

Citations

Mar 21, 2000·Journal of Cellular Biochemistry·M R SeghatoleslamiK A Knudsen
Jan 1, 1996·Developmental Genetics·A Buonanno, N Rosenthal
Jul 6, 2000·Developmental Dynamics : an Official Publication of the American Association of Anatomists·J WeisA Buonanno

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