Activity-dependent induction of slow myosin gene expression in isolated fast-twitch mouse muscle.
We demonstrate that direct electrical stimulation of isolated fast-twitch muscle in an organ culture system can induce expression of the slow myosin heavy chain (beta-MHC) gene, indicative of a phenotype transformation. Pairs of extensor digitorum longus (EDL) muscles were isolated from adult mice, incubated at resting length in separate chambers, and superfused with the same recirculated media One muscle was subjected to twitch stimulation (5-s trains of 5-Hz pulses at supramaximal voltage every minute), and force was recorded to assess function. The contralateral muscle was incubated without stimulation, to control for effects of the experimental preparation. Both muscle were rapidly frozen for RNA purification and oligo(dT)-primed reverse transcription; serial studies were carried out to 36 h. Polymerase chain reaction was performed utilizing primers specific for cytoplasmic beta-actin (beta-actin), a constitutive marker, and beta-MHC, a gene that is either inactive or expressed at very low levels in control EDL. After 30 h of stimulation, beta-MHC was consistently detected at a level severalfold higher in stimulated EDL than in incubated control EDL when band intensities were normalized to those of beta-actin. These results show that signals or fiber-specific transformations reside within the muscle and that this shift begins rapidly after induction of continuous stimulation.
Department of Physiology, University of Washington, Seattle 98195, USA.
E R
ER
Barton-Davis
W A
WA
LaFramboise
M J
MJ
Kushmerick
D000818:Animals; D004558:Electric Stimulation; D015870:Gene Expression; D008297:Male; D051379:Mice; D009119:Muscle Contraction; D018482:Muscle, Skeletal / Q000502:physiology*; D018995:Myosin Heavy Chains / Q000235:genetics*; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000235:genetics
D012333:RNA, Messenger; D018995:Myosin Heavy Chains