We previously reported the relationship between alpha1-adrenoceptor-mediated contraction and phosphorylation of 20-kDa myosin light chain (LC20) in de-endothelialized rat caudal arterial smooth muscle at room temperature (Mita M, Walsh MP. Biochem J. 1997;327:669-674). We now describe the effect of increasing the temperature to 37 degrees C on this relationship. The EC50 value (76.6 +/- 18.2 nM) for cirazoline (alpha1-adrenergic agonist)-induced contraction of the strips at room temperature (23 degrees C) was significantly greater than that (14.5 +/- 1.9 nM) at 37 degrees C. The initial rate of the contraction to a sub-maximal concentration of cirazoline (0.3 microM) was similar at the two temperatures. However, cirazoline-induced maximal force at 37 degrees C was approximately 1.8 times that at room temperature. LC20 phosphorylation in response to cirazoline at room temperature and 37 degrees C closely matched the time courses of contraction, but values were not significantly different at the two temperatures: resting phosphorylation levels were 0.09 +/- 0.04 mol P(i)/mol LC20 at 37 degrees C and 0.22 +/- 0.06 mol P(i)/mol LC20 at room temperature; maximal cirazoline-stimulated LC20 phosphorylation levels were 0.58 +/- 0.08 mol P(i)/mol LC20 at room temperature and 0.49 +/- 0.05 mol P(i)/mol LC20 at 37 degrees C. We conclude, therefore, that the enhanced cirazoline-induced contraction at 37 degrees C is not due to increased LC20 phosphorylation.