QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: mol.104.002642v1 66/5/1265    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Shen, J. Articles by Sturek, M. Search for Related Content PubMed PubMed Citation Articles by Shen, J. Articles by Sturek, M.

Cloning, Up-Regulation, and Mitogenic Role of Porcine P2Y₂ Receptor in Coronary Artery Smooth Muscle Cells

Departments of Medical Pharmacology and Physiology (J.S., M.W., P.A.W., M.S.), Biochemistry (C.I.S., G.A.W.), and Internal Medicine (M.S.), and the Center for Diabetes and Cardiovascular Health (J.S., M.W., P.A.W., M.S.), University of Missouri-Columbia, School of Medicine, Columbia, Missouri

Previous work has shown up-regulation of a UTP-sensitive P2Y receptor in porcine coronary smooth muscle cells (CSMC) of organ-cultured arteries. However, the molecular identity and functional role of this putative receptor remained undefined. Here we report the cloning of the cDNA for this receptor that encodes an open reading frame for a protein of 373 amino acids with the highest homology to the human P2Y₂ receptor (84%). Heterologous expression of this receptor in 1321N1 cells revealed a novel pharmacology in that UTP and ITP were full agonists and UTP was more potent and efficacious than ATP for increasing intracellular [Ca²⁺] and extracellular signal-regulated kinase phosphorylation. Stimulation of subcultured CSMC with UTP, ITP, or ATP induced a concentration-dependent increase in cellular DNA content, protein synthesis, cell number, and proliferating cell nuclear antigen expression, indicating a mitogenic role for P2Y₂ receptors. This was supported by the finding that the treatment of CSMC with antisense oligonucleotides to the cloned cDNA sequence significantly inhibited UTP- and ATP-induced DNA and protein synthesis. In addition, reverse transcription-polymerase chain reaction analysis showed that P2Y₂ receptor mRNA was dramatically increased in cells of organ-cultured arteries compared with freshly harvested arteries, whereas the P2Y₆ receptor mRNA level was unchanged, and the P2Y₄ receptor mRNA was undetectable. This P2Y₂ subtype-specific up-regulation was confirmed in cells of coronary arteries stented in vivo. In conclusion, we have cloned the porcine P2Y₂ receptor with novel pharmacology and demonstrated that this receptor is up-regulated in CSMC of in vitro organ cultures or in vivo stented coronary arteries to mediate the mitogenic effects of nucleotides.

Received for publication May 14, 2004.

Accepted for publication July 26, 2004.

Address correspondence to: Dr. Michael Sturek, Professor and Chair, Department of Cellular and Integrative Physiology, Indiana University, School of Medicine, 635 Barnhill Drive, MS 309, Indianapolis, IN 46202-5120. E-mail: msturek{at}iupui.edu

This article has been cited by other articles:

				J. Shen and P. E. DiCorleto ADP Stimulates Human Endothelial Cell Migration via P2Y1 Nucleotide Receptor-Mediated Mitogen-Activated Protein Kinase Pathways Circ. Res., February 29, 2008; 102(4): 448 - 456. [Abstract] [Full Text] [PDF]

				S. Jalvy, M.-A. Renault, L. Lam Shang Leen, I. Belloc, A. Reynaud, A.-P. Gadeau, and C. Desgranges CREB Mediates UTP-Directed Arterial Smooth Muscle Cell Migration and Expression of the Chemotactic Protein Osteopontin via Its Interaction with Activator Protein-1 Sites Circ. Res., May 11, 2007; 100(9): 1292 - 1299. [Abstract] [Full Text] [PDF]

				M. P. Abbracchio, G. Burnstock, J.-M. Boeynaems, E. A. Barnard, J. L. Boyer, C. Kennedy, G. E. Knight, M. Fumagalli, C. Gachet, K. A. Jacobson, et al. International Union of Pharmacology LVIII: Update on the P2Y G Protein-Coupled Nucleotide Receptors: From Molecular Mechanisms and Pathophysiology to Therapy Pharmacol. Rev., September 1, 2006; 58(3): 281 - 341. [Abstract] [Full Text] [PDF]

				J. Shen, S. P. Halenda, M. Sturek, and P. A. Wilden Cell-Signaling Evidence for Adenosine Stimulation of Coronary Smooth Muscle Proliferation via the A1 Adenosine Receptor Circ. Res., September 16, 2005; 97(6): 574 - 582. [Abstract] [Full Text] [PDF]

				J. Shen, S. P. Halenda, M. Sturek, and P. A. Wilden Novel Mitogenic Effect of Adenosine on Coronary Artery Smooth Muscle Cells: Role for the A1 Adenosine Receptor Circ. Res., May 13, 2005; 96(9): 982 - 990. [Abstract] [Full Text] [PDF]