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Molecular Pharmacology

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Rapid CommunicationAccelerated Communication

Selective Enhancement of β-Adrenergic Receptor Signaling by Overexpression of Adenylyl Cyclase Type 6: Colocalization of Receptor and Adenylyl Cyclase in Caveolae of Cardiac Myocytes

Rennolds S. Ostrom, Jonathan D. Violin, Scott Coleman and Paul A. Insel
Molecular Pharmacology May 2000, 57 (5) 1075-1079;
Rennolds S. Ostrom
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Jonathan D. Violin
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Scott Coleman
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Paul A. Insel
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Abstract

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), H2agonist, glucagon, and A2 agonist were not increased. Studies to test whether the selective enhancement in β-adrenergic receptor (AR) response might result from inhibition of AC6 by Gαi and Gβγ 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 β1-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 β-AR-stimulated cAMP formation and that key components of β-AR-mediated activation of AC exist in caveolae of cardiac myocytes, providing a means by which β-AR response is selectively enhanced by increasing AC6 expression.

Footnotes

  • Send reprint requests to: Paul A. Insel, M.D., Dept. of Pharmacology, 0636, University of California, San Diego, La Jolla, CA. E-mail: pinsel{at}ucsd.edu

  • ↵1 These authors contributed equally to this work.

  • This work was supported by research and training grants from the National Institutes of Health.

  • Abbreviations:
    AC
    adenylyl cyclase
    AC5 and AC6
    AC types 5 and 6
    PGE2
    prostaglandin E2
    HA
    hemagglutinin
    AR
    adrenergic receptor
    GFP
    green fluorescent protein
    • Received December 16, 1999.
    • Accepted February 7, 2000.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 57 (5)
Molecular Pharmacology
Vol. 57, Issue 5
1 May 2000
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Rapid CommunicationAccelerated Communication

Selective Enhancement of β-Adrenergic Receptor Signaling by Overexpression of Adenylyl Cyclase Type 6: Colocalization of Receptor and Adenylyl Cyclase in Caveolae of Cardiac Myocytes

Rennolds S. Ostrom, Jonathan D. Violin, Scott Coleman and Paul A. Insel
Molecular Pharmacology May 1, 2000, 57 (5) 1075-1079;

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Rapid CommunicationAccelerated Communication

Selective Enhancement of β-Adrenergic Receptor Signaling by Overexpression of Adenylyl Cyclase Type 6: Colocalization of Receptor and Adenylyl Cyclase in Caveolae of Cardiac Myocytes

Rennolds S. Ostrom, Jonathan D. Violin, Scott Coleman and Paul A. Insel
Molecular Pharmacology May 1, 2000, 57 (5) 1075-1079;
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