Expression of alpha -Adrenoceptor Subtypes by Smooth Muscle Cells and Adventitial Fibroblasts in Rat Aorta and in Cell Culture

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Vol. 298, Issue 2, 441-452, August 2001

Expression of $alpha$ -Adrenoceptor Subtypes by Smooth Muscle Cells and Adventitial Fibroblasts in Rat Aorta and in Cell Culture

James E. Faber, Nengyu Yang and Xiaohua Xin

Department of Cell and Molecular Physiology, School of Medicine, University of North Carolina, Chapel Hill, North Carolina

Previous radioligand binding reports of vascular $alpha$ -adrenoceptor (AR) density have been limited to total $alpha$ ₁- or $alpha$ ₂-ARs. Studiesusing whole blood vessel homogenates have not differentiated amongreceptor or mRNA expression by medial smooth muscle cells (SMCs)versus adventitial fibroblasts (AFBs). Therefore, we used quantitativereverse transcription-polymerase chain reaction and radioligandbinding to measure $alpha$ -AR subtypes in media, adventitia, and culturedSMCs and AFBs from rat aorta. Both media and adventitia expressed $alpha$ _1A-, $alpha$ _1B-, $alpha$ _1D-, and $alpha$ _2D-AR mRNAs, but in markedly differentabundances. Total $alpha$ ₁-AR density was the same for media and adventitia(B_max = 101 ± 10 versus 96 ± 16 fmol/mg of protein). However,densities for $alpha$ _1A-, $alpha$ _1B-, and $alpha$ _1D-AR subtypes in media were 19± 2, 26 ± 4, and 55 ± 2%, and in adventitia were 44 ± 3, 37 ±5, and 19 ± 2%. No $alpha$ _2B- or $alpha$ _2C-AR transcripts were detected ineither layer or in cultured SMCs or AFBs. Total $alpha$ ₁-AR densitiesin cultured SMCs and AFBs (B_max = 111 ± 4 and 48 ± 6 fmol/mg ofprotein, respectively) were similar to media and adventitia, with $alpha$ _1B- and $alpha$ _1D-AR transcript levels and receptors largely sustained.However, $alpha$ _1A- and $alpha$ _2D-AR expression in cultured SMCs and AFBswas strongly reduced, compared with media and adventitia, an effectnot prevented by 30 different culture conditions. Like SMCs, exposureof AFBs to norepinephrine induced protein synthesis and proliferationof AFBs. This is the first study to quantitate $alpha$ -AR subtype expressionin media and adventitia and in cultured SMCs and AFBs. In addition,we report the intriguing finding that AFBs express $alpha$ ₁-ARs in similarabundance as medial SMCs and that norepinephrine induced themtoproliferate.

0022-3565/01/2982-0441$03.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 2001 by The American Society for Pharmacology and Experimental Therapeutics

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				T. Kobayashi, T. Matsumoto, K. Ooishi, and K. Kamata Differential expression of {alpha}2D-adrenoceptor and eNOS in aortas from early and later stages of diabetes in Goto-Kakizaki rats Am J Physiol Heart Circ Physiol, July 1, 2004; 287(1): H135 - H148. [Abstract] [Full Text] [PDF]

				M. A. Chotani, S. Mitra, B. Y. Su, S. Flavahan, A. H. Eid, K. R. Clark, C. R. Montague, H. Paris, D. E. Handy, and N. A. Flavahan Regulation of {alpha}2-adrenoceptors in human vascular smooth muscle cells Am J Physiol Heart Circ Physiol, January 1, 2004; 286(1): H59 - H67. [Abstract] [Full Text] [PDF]

				S. A Ross, B. R Rorabaugh, D. Chalothorn, J. Yun, P. J Gonzalez-Cabrera, D. F McCune, M. T Piascik, and D. M Perez The {alpha}1B-adrenergic receptor decreases the inotropic response in the mouse Langendorff heart model Cardiovasc Res, December 1, 2003; 60(3): 598 - 607. [Abstract] [Full Text] [PDF]

				J. J. Carrillo, J. Pediani, and G. Milligan Dimers of Class A G Protein-coupled Receptors Function via Agonist-mediated Trans-activation of Associated G Proteins J. Biol. Chem., October 24, 2003; 278(43): 42578 - 42587. [Abstract] [Full Text] [PDF]

				P. J. Gonzalez-Cabrera, R. J. Gaivin, J. Yun, S. A. Ross, R. S. Papay, D. F. McCune, B. R. Rorabaugh, and D. M. Perez Genetic Profiling of alpha 1-Adrenergic Receptor Subtypes by Oligonucleotide Microarrays: Coupling to Interleukin-6 Secretion but Differences in STAT3 Phosphorylation and gp-130 Mol. Pharmacol., May 1, 2003; 63(5): 1104 - 1116. [Abstract] [Full Text] [PDF]

				S. Kalyankrishna and K. U. Malik Norepinephrine-Induced Stimulation of p38 Mitogen-Activated Protein Kinase Is Mediated by Arachidonic Acid Metabolites Generated by Activation of Cytosolic Phospholipase A2 in Vascular Smooth Muscle Cells J. Pharmacol. Exp. Ther., February 1, 2003; 304(2): 761 - 772. [Abstract] [Full Text] [PDF]

Expression of -Adrenoceptor Subtypes by Smooth Muscle Cells and Adventitial Fibroblasts in Rat Aorta and in Cell Culture

Expression of $alpha$ -Adrenoceptor Subtypes by Smooth Muscle Cells and Adventitial Fibroblasts in Rat Aorta and in Cell Culture

				J. C. Teeters, C. Erami, H. Zhang, and J. E. Faber Systemic alpha 1A-adrenoceptor antagonist inhibits neointimal growth after balloon injury of rat carotid artery Am J Physiol Heart Circ Physiol, January 1, 2003; 284(1): H385 - H392. [Abstract] [Full Text] [PDF]

				C. Hague, P. J. Gonzalez-Cabrera, W. B. Jeffries, and P. W. Abel Relationship between alpha 1-Adrenergic Receptor-Induced Contraction and Extracellular Signal-Regulated Kinase Activation in the Bovine Inferior Alveolar Artery J. Pharmacol. Exp. Ther., October 1, 2002; 303(1): 403 - 411. [Abstract] [Full Text] [PDF]

				C. Erami, H. Zhang, J. G. Ho, D. M. French, and J. E. Faber alpha 1-Adrenoceptor stimulation directly induces growth of vascular wall in vivo Am J Physiol Heart Circ Physiol, October 1, 2002; 283(4): H1577 - H1587. [Abstract] [Full Text] [PDF]

				H. Zhang, C. S. Facemire, A. J. Banes, and J. E. Faber Different alpha -adrenoceptors mediate migration of vascular smooth muscle cells and adventitial fibroblasts in vitro Am J Physiol Heart Circ Physiol, June 1, 2002; 282(6): H2364 - H2370. [Abstract] [Full Text] [PDF]

				H. Zhang and J. E. Faber Trophic Effect of Norepinephrine on Arterial Intima-Media and Adventitia Is Augmented by Injury and Mediated by Different {alpha}1-Adrenoceptor Subtypes Circ. Res., October 26, 2001; 89(9): 815 - 822. [Abstract] [Full Text] [PDF]