Enhanced responsiveness of the myocardial beta-adrenoceptor-adenylate cyclase system in the perfused rat heart (I).
Crude myocardial sarcolemmal membrane fractions were prepared from rat hearts subjected to total global ischemia with and without normoxic reperfusion, or global anoxic (N2) perfusion with and without normoxic reperfusion. The direct effects on beta-adrenoceptor number, G-protein levels and stimulation of the adenylate cyclase (AC) complex were assessed. In terms of AC activation, ischemia led to a marked increase (4-fold) in sensitivity to terbutaline (beta2-agonist) and phorbol ester (tetradecanoyl phorbol acetate = TPA) stimulation, whereas the dobutamine (beta1) responsiveness and Gpp(NH)p activation through G(s)alpha/G(i2)alpha remained unaltered. However, forskolin-elicited holoenzyme activity fell markedly during normoxic reperfusion. Ischemia did not change the beta1-adrenoceptor number, while beta2-receptor population increased by approximately 45%. Western blots of myocardial G(s)A and G(i2)alpha contents revealed that ischemia selectively diminished G(i2)alpha levels only by some 50-70%. Contrastingly, anoxia selectively increased the AC sensitivity (2-fold) to beta1-adrenergic stimulation. As subsequent to ischemia, anoxia also increased the sensitivity to TPA stimulation, however, only 2-fold. Gpp(NH)p activation was unchanged, while forskolin-enhanced activity gradually declined, also during ensuing normoxic reperfusion. Anoxia brought about a 75% enhancement in beta1-receptor number, while beta2-receptors remained unaffected. However, altered receptor number normalized on termination of normoxic reperfusion. Finally, anoxia led to a 50-60% decimation of myocardial G(i2)alpha levels, while G(s)alpha was only marginally reduced. Despite the fact that the ischemia and anoxia effectuated a similar deterioration of physiological heart parameters, myocardial contents of energy rich phosphate moieties and loss of G(i2)alpha, ischemia rendered the most profound increase in responsiveness of the sarcolemmal AC system.
Gordeladze
J O
JO
Institute for Surgical Research, The National Hospital, Rikshospitalet, University of Oslo, Norway.
Jynge
I
I
Borchgrevink
P C
PC
Sellevold
O F
OF
D000262:Adenylyl Cyclases / Q000187:drug effects / Q000378:metabolism*; D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000319:Adrenergic beta-Antagonists / Q000494:pharmacology; D000818:Animals; D002078:Butoxamine / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D006321:Heart / Q000187:drug effects / Q000503:physiopathology; D006352:Heart Ventricles / Q000378:metabolism; D000860:Hypoxia / Q000188:drug therapy / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D017202:Myocardial Ischemia / Q000378:metabolism* / Q000503:physiopathology; D009206:Myocardium / Q000378:metabolism*; D010477:Perfusion; D051381:Rats; D017208:Rats, Wistar; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D012508:Sarcolemma / Q000201:enzymology; D013726:Terbutaline / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000318:Adrenergic beta-Agonists; D000319:Adrenergic beta-Antagonists; D011943:Receptors, Adrenergic, beta; D002078:Butoxamine; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D013726:Terbutaline; D013755:Tetradecanoylphorbol Acetate