This study provides new information about the relative importance of different alpha(1)-adrenoceptors during norepinephrine (NE) activation in rat renal resistance vessels. In Sprague-Dawley rats, we measured renal blood flow (RBF) using electromagnetic flowmetry in vivo and the intracellular free calcium concentration ([Ca(2+)](i)) utilizing ratiometric photometry of fura 2 fluorescence in isolated afferent arterioles. Renal arterial bolus injection of NE produced a transient 46% decrease in RBF. In microdissected afferent arterioles, NE (1 microM) elicited an immediate square-shaped increase in [Ca(2+)](i), from 90 to 175 nM (P < 0.001). Chloroethylclonidine (CEC) (50 microM) had no chronic irreversible alkylating effect in vitro but exerted acute reversible blockade on norepinephrine (NE) responses both on [Ca(2+)](i) in vitro and on RBF in vivo. The RBF response was attenuated by approximately 50% by the putative alpha(1A)-adrenoceptor and alpha(1D)-adrenoceptor antagonists 5-methylurapidil (5-MU), and 8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4. 5]decane-7,9-dione dihydrochloride (BMY-7378) (12.5 and 62.5 microg/h), respectively. The in vitro [Ca(2+)](i) response to NE was blocked approximately 25% and 50% by 5-MU (100 nM and 1 microM). BMY-7378 (100 nM and 1 microM) attenuated the NE-induced response by approximately 40% and 100%. The degree of inhibition in vitro was similar to the in vivo experiments. In conclusion, 5-MU and BMY-7378 attenuated the NE-induced responses, although relatively high concentrations were required, suggesting involvement of both the alpha(1A)-adrenoceptor and alpha(1D)-adrenoceptor. Participation of the alpha(1B)-adrenoceptor is less likely, as we found no evidence for CEC-induced alkylation.