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Updated: Aug 29, 2026

Bladder Smooth Muscle Strip Contractility as a Method to Evaluate Lower Urinary Tract Pharmacology
Published on: August 18, 2014
Domain-dependent action of urokinase on smooth muscle cell responses
William J Tanski1, Allison J Fegley, Elisa Roztocil
1Department of Surgery, University of Rochester, Strong Memorial Hospital, Box SURG, 601 Elmwood Avenue, Rochester, NY 14642, USA. wtanski@rochester.rr.com
Background:
Single-chain urokinase-type plasminogen activator (sc-uPA) is one of the key serine proteases involved in modulating cellular and extracellular matrix responses during tissue remodeling. Sc-uPA is composed of three domains: aminoterminal fragment (ATF), kringle domain, and carboxyterminal fragment (CTF). sc-uPA is readily cleaved into these three domain fragments in vitro, each of which is biologically active; however, their roles in the microenvironment of the vessel wall are poorly understood.
Purpose:
The purpose of this study was to determine the role of each domain of sc-uPA on vascular smooth muscle cell (SMC) proliferation and migration.
Methods:
SMCs were cultured in vitro. Assays of DNA synthesis, cell proliferation, and migration were performed in response to sc-uPA, ATF, kringle, and CTF in the presence and absence of the plasmin inhibitors epsilon-aminocaproic acid (EACA) and aprotinin, the Galphai inhibitor pertussis toxin, and the mitogen-activated protein kinase 1 (the upstream regulator of the extracellular-signal regulated kinase [ERK]) inhibitor PD98059.
Results:
sc-uPA produced dose-dependent increases in DNA synthesis and cell proliferation. These responses were dependent on the CTF domain and were sensitive to plasmin inhibitors, pertussis toxin, and PD98059. Sc-uPA also induced SMC migration, which could be elicited by both ATF and kringle. Migration to sc-uPA, ATF, and kringle was both pertussis toxin and PD98059 sensitive, but importantly was plasmin-independent.
Conclusion:
sc-uPA induces SMC proliferation and migration, which are domain-dependent and mediated in part by Galphai-linked, ERK-dependent processes, while only the mitogenic response is protease dependent. These findings suggest that migration is linked to a G-protein coupled nonprotease receptor, while proliferation is associated with a G-protein coupled protease receptor.
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