Effects of epinephrine infusion on expression of calpastatin in porcine cardiac and skeletal muscle.
beta-Adrenergic agonists induce muscle hypertrophy in mammalian species and alter the extractable activity of calpain proteinase and its specific endogenous inhibitor calpastatin. The effects on skeletal and cardiac muscle calpastatin of continuously infusing a group of pigs for 7 days with the physiological agonist epinephrine (0.15 microg/kg/min) were examined and compared with a placebo group. Basal levels of extractable calpastatin activity were higher in cardiac than skeletal muscle and epinephrine infusion increased the extractable activity in both muscle types (P < 0.05). An anti-recombinant porcine calpastatin antiserum detected a 135-kDa band and a 145/135-kDa doublet on Western blots of skeletal and cardiac extracts, respectively. Epinephrine infusion increased the 135-kDa band in skeletal muscle (P < 0.05), while the ratio of 145/135 kDa in cardiac muscle was decreased (P < 0.05). From Northern blots, the patterns of calpastatin mRNA species were similar in the two muscle types, two major transcripts at 5.8 and 3. 2 kb in cardiac muscle, with equivalent bands in skeletal muscle of 5.4 and 2.8 kb. A faint 7.9-kb band was also detected in skeletal muscle. Epinephrine infusion had no effect on skeletal calpastatin mRNA but tended to increase the 5.8-kb mRNA in cardiac muscle (P = 0. 053). These data indicate a differential response of the two muscle types to mildly elevated plasma epinephrine concentration and the response to elevated epinephrine may be at the translational or posttranslational level. Therefore, this type of stimulus appears to be less effective at perturbing calpastatin gene transcription than certain orally administered synthetic beta-adrenergic agonists.
Parr
T
T
Division of Nutritional Biochemistry, University of Nottingham, Loughborough, Leicestershire, LE12 5RD, United Kingdom.
Sensky
P L
PL
Arnold
M K
MK
Bardsley
R G
RG
Buttery
P J
PJ
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D004837:Epinephrine / Q000008:administration & dosage / Q000097:blood / Q000494:pharmacology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D007262:Infusions, Intravenous; D008970:Molecular Weight; D018482:Muscle, Skeletal / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D013552:Swine; D014158:Transcription, Genetic / Q000187:drug effects
D002135:Calcium-Binding Proteins; D012333:RNA, Messenger; C033668:calpastatin; D004837:Epinephrine