Mechanical activity in heart regulates translation of alpha-myosin heavy chain mRNA but not its localization.
Mechanical inactivity depresses protein expression in cardiac muscle tissue and results in atrophy. We explore the mechanical transduction mechanism in spontaneously beating neonatal rat cardiomyocytes expressing the alpha-myosin heavy chain (alpha-MyHC) isoform by interfering with cross-bridge function [2,3-butanedione monoxime (BDM), 7.5 mM] without affecting cell calcium. The polysome content and alpha-MyHC mRNA levels in fractions from a sucrose gradient were analyzed. BDM treatment blocked translation at initiation (162 +/- 12% in the nonpolysomal RNA fraction and 43 +/- 6% in the polysomal fraction, relative to control as 100%; P < 0.05). There was an increase in alpha-MyHC mRNA from the nonpolysomal fraction (120.5 +/- 7.7%; P < 0.05 compared with control) with no significant change in the heavy polysomes. In situ hybridization of alpha-MyHC mRNA was used to estimate message abundance as a function of the distance from the nucleus. The mRNA was dispersed through the cytoplasm in spontaneously beating cells as well as in BDM-treated cells (no significant difference). We conclude that direct inhibition of contractile machinery, but not calcium, regulates initiation of alpha-MyHC mRNA translation. However, calcium, not pure mechanical signals, appears to be important for message localization.
Nikcevic
G
G
Department of Physiology and Biophysics, College of Medicine, University of Illinois at Chicago, Chicago, Illinois 60612, USA.
Heidkamp
M C
MC
Perhonen
M
M
Russell
B
B
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D003931:Diacetyl / Q000031:analogs & derivatives / Q000494:pharmacology; D006321:Heart / Q000187:drug effects / Q000502:physiology*; D009206:Myocardium / Q000378:metabolism / Q000648:ultrastructure; D009210:Myofibrils / Q000648:ultrastructure; D018995:Myosin Heavy Chains / Q000235:genetics*; D014176:Protein Biosynthesis / Q000502:physiology*; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000502:physiology*; D014018:Tissue Distribution / Q000502:physiology
D012333:RNA, Messenger; C004717:diacetylmonoxime; D012313:RNA; D018995:Myosin Heavy Chains; D003931:Diacetyl