OBJECTIVE: To quantify mRNA levels of Rho A, and rho-associated coil- forming kinases I and II (ROCK I and ROCK II) in human pregnant (before and after labor onset) and nonpregnant myometrium, and to investigate protein expression of Rho A, ROCK I, and ROCK II in these three tissue types.METHODS: Real-time fluorescence reverse transcriptase-polymerase chain reaction RT-PCR) using primers for Rho A, ROCK I, and ROCK II was performed on total RNA isolated from the three tissue types under investigation. Western blot analysis using antibodies specific to Rho A, ROCK I, and ROCK II was performed on protein isolated from the three tissue types.RESULTS: Real-time fluorescence RT-PCR, using primers for Rho A, ROCK I, and ROCK II, revealed Rho A mRNA expression was significantly greater in human pregnant myometrium after labor onset, in comparison to pregnant myometrium before labor onset (P .05). With Western blot analysis, using antibodies specific to Rho A, ROCK I, and ROCK II, Rho A protein levels were significantly lower in pregnant (before and after labor onset) in comparison to nonpregnant myometrium (P .05). No signal for ROCK II was detected in myometrial tissue.CONCLUSIONS: These results outline the presence of the Rho A/Rho kinase system in modulating contractility of human myometrium. Total Rho A protein expression is down-regulated in the third trimester of pregnancy while up-regulation of Rho A mRNA occurs with labor onset. Copyright (C) 2005 by the Society for Gynecologic Investigation.