Potentiation of beta-adrenergic signaling by adenoviral-mediated gene transfer in adult rabbit ventricular myocytes.
Abstract
Our laboratory has been testing the hypothesis that genetic modulation of the beta-adrenergic
signaling cascade can enhance cardiac function. We have previously shown that transgenic
mice with cardiac overexpression of either the human beta2-adrenergic receptor (beta2AR)
or an inhibitor of the beta-adrenergic receptor kinase (betaARK), an enzyme that phosphorylates
and uncouples agonist-bound receptors, have increased myocardial inotropy. We now
have created recombinant adenoviruses encoding either the beta2AR (Adeno-beta2AR)
or a peptide betaARK inhibitor (consisting of the carboxyl terminus of betaARK1, Adeno-betaARKct)
and tested their ability to potentiate beta-adrenergic signaling in cultured adult
rabbit ventricular myocytes. As assessed by radioligand binding, Adeno-beta2AR infection
led to approximately 20-fold overexpression of beta-adrenergic receptors. Protein
immunoblots demonstrated the presence of the Adeno-betaARKct transgene. Both transgenes
significantly increased isoproterenol-stimulated cAMP as compared to myocytes infected
with an adenovirus encoding beta-galactosidase (Adeno-betaGal) but did not affect
the sarcolemmal adenylyl cyclase response to Forskolin or NaF. beta-Adrenergic agonist-induced
desensitization was significantly inhibited in Adeno-betaARKct-infected myocytes (16+/-2%)
as compared to Adeno-betaGal-infected myocytes (37+/-1%, P < 0.001). We conclude that
recombinant adenoviral gene transfer of the beta2AR or an inhibitor of betaARK-mediated
desensitization can potentiate beta-adrenergic signaling.
Type
Journal articleSubject
AdenoviridaeAdenylyl Cyclases
Adrenergic beta-Agonists
Animals
Cell Survival
Cells, Cultured
Cyclic AMP
Gene Transfer Techniques
Genetic Vectors
Heart Ventricles
Humans
Isoproterenol
Male
Rabbits
Receptors, Adrenergic, beta-2
Sarcolemma
Signal Transduction
Transgenes
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https://hdl.handle.net/10161/7831Published Version (Please cite this version)
10.1172/JCI119157Publication Info
Drazner, MH; Peppel, KC; Dyer, S; Grant, AO; Koch, WJ; & Lefkowitz, RJ (1997). Potentiation of beta-adrenergic signaling by adenoviral-mediated gene transfer in
adult rabbit ventricular myocytes. J Clin Invest, 99(2). pp. 288-296. 10.1172/JCI119157. Retrieved from https://hdl.handle.net/10161/7831.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Augustus Oliver Grant
Professor of Medicine
This laboratory seeks to understand the molecular events that control excitation
in cardiac muscle and the mechanisms by which excitation is modified by drugs. The
experimental models include the isolated cardiac myocyte, the frog oocyte and mammalian
cells expressing mRNA transcripts of wild-type and mutant channels. The experimental
techniques include DNA and RNA manipulation, whole cell voltage clamping and the extracellular
patch clamping. The current focus of the laboratory i
Robert J. Lefkowitz
The Chancellor's Distinguished Professor of Medicine
Dr. Lefkowitz’s memoir, A Funny Thing Happened on the Way to Stockholm, recounts his
early career as a cardiologist and his transition to biochemistry, which led to his
Nobel Prize win.
Robert J. Lefkowitz, M.D. is James B. Duke Professor of Medicine and Professor of
Biochemistry and Chemistry at the Duke University Medical Center. He has been an Investigator
of the
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