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(Circulation Research. 2000;86:43.)
© 2000 American Heart Association, Inc.

Cellular Biology

Hybrid Transgenic Mice Reveal In Vivo Specificity of G Protein–Coupled Receptor Kinases in the Heart

Andrea D. Eckhart, Sandra J. Duncan, Raymond B. Penn, Jeffrey L. Benovic, Robert J. Lefkowitz, Walter J. Koch

From the Department of Surgery (A.D.E., S.J.D., W.J.K.) and the Department of Medicine and Biochemistry and The Howard Hughes Medical Institute (R.J.L.), Duke University Medical Center, Durham, NC, and the Department of Microbiology and Immunology (R.B.P., J.L.B.), Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, Pa.

Correspondence to Walter J. Koch, Department of Surgery, Box 2606, Room 472, MSRB, Duke University Medical Center, Durham, NC 27710. E-mail koch0002{at}mc.duke.edu

Abstract—G protein–coupled receptor kinases (GRKs) phosphorylate activated G protein-coupled receptors, including _1B-adrenergic receptors (ARs), resulting in desensitization. In vivo analysis of GRK substrate selectivity has been limited. Therefore, we generated hybrid transgenic mice with myocardium-targeted overexpression of 1 of 3 GRKs expressed in the heart (GRK2 [commonly known as the ß-AR kinase 1], GRK3, or GRK5) with concomitant cardiac expression of a constitutively activated mutant (CAM) or wild-type _1BAR. Transgenic mice with cardiac CAM_1BAR overexpression had enhanced myocardial ₁AR signaling and elevated heart-to-body weight ratios with ventricular atrial natriuretic factor expression denoting myocardial hypertrophy. Transgenic mouse hearts overexpressing only GRK2, GRK3, or GRK5 had no hypertrophy. In hybrid transgenic mice, enhanced in vivo signaling through CAM_1BARs, as measured by myocardial diacylglycerol content, was attenuated by concomitant overexpression of GRK3 but not GRK2 or GRK5. CAM_1BAR-induced hypertrophy and ventricular atrial natriuretic factor expression were significantly attenuated with either concurrent GRK3 or GRK5 overexpression. Similar GRK selectivity was seen in hybrid transgenic mice with wild-type _1BAR overexpression concurrently with a GRK. GRK2 overexpression was without effect on any in vivo CAM or wild-type _1BAR cardiac phenotype, which is in contrast to previously reported in vitro findings. Furthermore, endogenous myocardial ₁AR mitogen-activated protein kinase signaling in single-GRK transgenic mice also exhibited selectivity, as GRK3 and GRK5 desensitized in vivo ₁AR mitogen–activated protein kinase responses that were unaffected by GRK2 overexpression. Thus, these results demonstrate that GRKs differentially interact with _1BARs in vivo such that GRK3 desensitizes all _1BAR signaling, whereas GRK5 has partial effects and, most interestingly, GRK2 has no effect on in vivo _1BAR signaling in the heart.

Key Words: desensitization • adrenergic receptors, ₁ • protein-coupled receptor kinase • myocardial biology • myocardial hypertrophy

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