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INFLAMMATION, IMMUNOPHARMACOLOGY, AND ASTHMA

Evaluation of Urothelial Stretch-Induced Cyclooxygenase-2 Expression in Novel Human Cell Culture and Porcine in Vivo Ureteral Obstruction Models

The Uropharmacology and Endourology Research Laboratory, Departments of Pharmaceutical Sciences (T.J.J., W.S.M.) and Surgery-Division of Urology (T.J.J., D.E.B., S.Y.N.), University of Wisconsin Schools of Pharmacy and Medicine, Madison, Wisconsin

Obstruction and stretch induce cyclooxygenase (COX)-2 expression and prostanoid synthesis in urinary tissues, causing pain, inflammation, hypercontractility, and cell proliferation. Our objective was to characterize acute COX-2 induction during in vivo ureteral obstruction, establish a cell culture model of urothelial stretch-induced COX-2 expression, and evaluate whether mechanotransduction could alter transcriptional and post-transcriptional regulation of COX-2. We performed laparoscopic unilateral ureteral ligation in pigs and allowed progression for 1, 2, 6, 24, or 48 h. We evaluated COX-2 expression with reverse transcriptase (RT)-polymerase chain reaction (PCR) and immunoblotting. We cultured primary human urothelial cells on stretch plates, applied stretch for up to 48 h, and measured COX-2 expression by RT-PCR and immunoblotting, transcription with run-on assays, and mRNA stability with actinomycin mRNA decay assays. In vivo ureteral obstruction induced COX-2 expression 4-fold within 6 h, maintaining induction for 24 h. In cell culture, stretch induced COX-2 steady-state mRNA and protein within the first 3 h of stretch, maintaining this induction for over 6 h. Three hours of stretch doubled COX-2 transcription relative to unstretched controls and increased COX-2 mRNA half-life 3-fold. This is the first report to characterize in vivo temporal stretch-induced COX-2 expression in the urothelium and establish a primary urothelial cell culture model for the study of stretch-induced COX-2 mechanisms. This is also the first report to identify alterations in steady-state COX-2 mRNA having components of both transcriptional and post-transcriptional regulation of stretch-regulated COX-2. Future elucidation of COX-2 signaling may identify novel therapeutic targets for treating stretch and distension of urinary tissues.

Address correspondence to: Dr. Travis J. Jerde, University of Wisconsin Medical School; Department of Surgery, Division of Urology, K6-561 Clinical Science Center, 600 Highland Avenue, Madison, WI 53792. E-mail: jerde{at}surgery.wisc.edu

This article has been cited by other articles:

				T. J. Jerde, W. S. Mellon, D. E. Bjorling, C. M. Checura, K. Owusu-Ofori, J. J. Parrish, and S. Y. Nakada Stretch Induction of Cyclooxygenase-2 Expression in Human Urothelial Cells Is Calcium- and Protein Kinase C {zeta}-Dependent Mol. Pharmacol., January 1, 2008; 73(1): 18 - 26. [Abstract] [Full Text] [PDF]