Selective enhancement of beta-adrenergic receptor signaling by overexpression of adenylyl cyclase type 6: colocalization of receptor and adenylyl cyclase in caveolae of cardiac myocytes.
We investigated the effect of adenovirally mediated overexpression of adenylyl cyclase type 6 (AC6), a major form of AC expressed in mammalian heart, on G protein-coupled receptor regulation of cAMP production in neonatal rat ventricular myocytes. Following gene transfer of AC6, isoproterenol- and forskolin-stimulated increases in cAMP were markedly enhanced, whereas basal levels of cAMP and responses to several other agonists that stimulate cAMP formation, e. g., prostaglandin E(2) (PGE(2)), H(2) agonist, glucagon, and A(2) agonist were not increased. Studies to test whether the selective enhancement in beta-adrenergic receptor (AR) response might result from inhibition of AC6 by Galpha(i) and Gbetagamma indicated that pertussis toxin-sensitive inhibition by the muscarinic cholinergic agonist carbachol was unaltered in myocytes overexpressing AC6. Pertussis toxin treatment failed to reveal an enhancement by AC6 overexpression of basal or PGE(2)-stimulated cAMP. Immunoblot analysis of membrane fractions indicated that beta(1)-AR and AC6 are expressed in fractions enriched in caveolin-3 and morphologic caveolae. The data suggest that loss of G(i)-mediated inhibition is not the mechanism for enhancement of beta-AR-stimulated cAMP formation and that key components of beta-AR-mediated activation of AC exist in caveolae of cardiac myocytes, providing a means by which beta-AR response is selectively enhanced by increasing AC6 expression.
Ostrom
R S
RS
Department of Pharmacology, School of Medicine, University of California, San Diego, La Jolla, CA, USA.
Violin
J D
JD
Coleman
S
S
Insel
P A
PA
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism*; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism; D015151:Immunoblotting; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism*; D051381:Rats; D011943:Receptors, Adrenergic, beta / Q000378:metabolism*; D012508:Sarcolemma / Q000378:metabolism; D015398:Signal Transduction
D011943:Receptors, Adrenergic, beta; D000242:Cyclic AMP; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D019205:GTP-Binding Protein alpha Subunits, Gs; D000262:Adenylyl Cyclases; C121100:adenylyl cyclase 6