Reduction of caveolin-3 expression does not inhibit stretch-induced phosphorylation of ERK2 in skeletal muscle myotubes.
Mechanotransduction is critical to the maintenance and growth of skeletal muscle, but the mechanism by which cellular deformations are converted to biochemical signals remains unclear. Among the earliest and most ubiquitous responses to mechanical stimulation is the phosphorylation and activation of mitogen-activated protein kinases, in particular ERK2. Caveolin-3 (CAV-3) binds ERK2 and its upstream activators in inactive states on the caveolae of resting muscle. Caveolae are deformed by stretch, and it was hypothesized that this deformation might disrupt the CAV-3-dependent inhibition of ERK2 to affect stretch-induced activation. Stretch-induced phosphorylation of ERK2 in myotubes was both amplitude and velocity dependent, consistent with a viscoelastic mechanism, such as deformation of caveolae. Chemical disruption of caveolae by cholesterol depletion increased ERK2 activation by up to 176%. Small interfering RNA oligomers were then used to knock down expression of CAV-3 in cultured myotubes before mechanical stimulation, with the expectation that reducing CAV-3 expression would eliminate the stretch-induced activation of ERK2. Knockdown reduced CAV-3 protein content by 55% but did not significantly alter the stretch-induced increase in ERK2 phosphorylation, suggesting that CAV-3 is not an essential element of the mechanotransduction pathway, although the limited extent of knockdown limits the strength of this conclusion.
Bellott
Anne Claire
AC
School of Applied Physiology, Georgia Institute of Technology, 281 Ferst Dr., Atlanta, GA 30332-0356, USA.
Patel
Kalpit C
KC
Burkholder
Thomas J
TJ
D000818:Animals; D051244:Caveolin 3; D022461:Caveolins / Q000172:deficiency*; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D018485:Muscle Fibers, Skeletal / Q000502:physiology*; D018482:Muscle, Skeletal / Q000502:physiology*; D010766:Phosphorylation; D010812:Physical Stimulation / Q000379:methods
C495548:Cav3 protein, mouse; D051244:Caveolin 3; D022461:Caveolins; D002784:Cholesterol; D019950:Mitogen-Activated Protein Kinase 1