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Vol. 57, Issue 5, 1075-1079, May 2000
-Adrenergic Receptor Signaling by
Overexpression of Adenylyl Cyclase Type 6: Colocalization of Receptor
and Adenylyl Cyclase in Caveolae of Cardiac Myocytes
Department of Pharmacology, School of Medicine, University of
California, San Diego, La Jolla, California
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 E2 (PGE2), H2
agonist, glucagon, and A2 agonist were not increased.
Studies to test whether the selective enhancement in 
-adrenergic
receptor (AR) response might result from inhibition of AC6 by
G
i and G
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
PGE2-stimulated cAMP. Immunoblot analysis of membrane fractions indicated that 1-AR and AC6 are expressed in
fractions enriched in caveolin-3 and morphologic caveolae. The data
suggest that loss of Gi-mediated inhibition is not the
mechanism for enhancement of -AR-stimulated cAMP formation and that
key components of -AR-mediated activation of AC exist in caveolae of
cardiac myocytes, providing a means by which -AR response is
selectively enhanced by increasing AC6 expression.
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