Novel in vitro system for functional assessment of oxytocin action

Departments of 1 Medicine, 2 Pharmacology and Therapeutics, and 3 Obstetrics and Gynecology, McGill University, Montreal, Quebec, Canada Submitted 2 November 2005 ; accepted in final form 28 August 2006 One of the classical biological actions mediated by the posterior pituitary hormone oxytocin (OT) is contraction of the uterus at parturition. Moreover, premature activation of the OT system is thought to contribute to preterm labor, a major clinical problem in obstetrical practice. However, the molecular mechanisms linking activation of the OT receptor (OTR) to myometrial contractions are not fully understood. Here, we describe an in vitro system that should serve as a useful tool to study this question at a cellular level. The system consists of a collagen lattice contraction assay and two different human myometrial cell lines: a cell clone from a telomerase-immortalized human myometrial cell population (hTERT-C3) as well as a cell line derived from a primary culture of human myometrial cells (M11). Using this approach, we observed that 1 nM OT promoted an almost maximal effect on cell contraction in both cell lines tested. Furthermore, this dose-dependent, OT-induced contraction was antagonized by the specific OTR antagonist d(CH 2 ) 5 [Tyr(Me) 2 ,Thr 4 ,Tyr-NH 2 9 ]OVT as well as the clinically used antagonist atosiban. This cell line-based contraction assay enables the application of molecular tools aimed at suppressing or overexpressing specific genes. It is also amenable to high-throughput testing approaches. Therefore, this system represents a powerful and improved experimental model that should facilitate the study of the molecular signal transduction pathways involved in the uterotonic actions of OT. oxytocin receptor; myometrium; parturition; myometrial cell lines; mitogen-activated protein kinase; extracellular signal-regulated kinase 1/2 Address for reprint requests and other correspondence: H. H. Zingg, Dept. of Pharmacology and Therapeutics, McGill University, 3655 Promenade Sir-William-Osler, Montreal, QC, H3G 1Y6, Canada (e-mail: hans.zingg{at}mcgill.ca )