Behavioral experiments examined the roles of substance P (SP) and cholecystokinin (CCK) in male rat copulatory behavior. Male copulatory behavior was recorded subsequent to injections of different doses of CCK and SP into the medial preoptic-anterior-hypothalamic area (MPOA-AH), caudate/putamen (CP), or the lateral ventricles (LV) in sexually experienced male rats. In the first experiment, three different doses of SP (10, 100, and 200 ng/cannula) injected bilaterally into the MPOA-AH produced marked changes in several components of male copulatory behavior. Latencies were most affected. All three doses significantly shortened the interval to initiate copulation, and the 10 and 100 ng, but not 200 ng dose also significantly reduced ejaculation latencies. Injections of 10 ng of SP into the CP did not affect sexual behavior, while injections into the LV produced changes different from those of MPOA-AH injections. These data argue for some degree of site specificity of the effects of the MPOA-AH injections. Bilateral injections of 10 ng of SP into the MPOA-AH, were incapable of inducing copulatory behavior in castrated rats deprived of testosterone. Injections of an undiluted SP antiserum (2 microliters/cannula) into the MPOA-AH produced a dramatic impairment of male copulatory behavior. These injections significantly lengthened amount, intromission, and ejaculation latencies, while having no effect on the number of mounts or intromissions prior to ejaculation. In contrast, bilateral injections of CCK-8 (10, 100, and 200 ng/cannula) into the MPOA-AH failed to affect any parameter of male copulatory behavior.(ABSTRACT TRUNCATED AT 250 WORDS)