The purpose of these studies was to ascertain which parameters of pulsatile gonadotropin secretion are regulated by endogenous inhibin in the intact diestrous female rat. This was determined by examining the changes in the secretion parameters of FSH and LH that resulted from immunoneutralizing endogenous inhibin in diestrous I female rats. Passive immunoneutralization of endogenous inhibin was achieved using specific, high titer ovine antiserum generated against the alpha-subunit of the recently described inhibin molecule. The optimal times after inhibin immunoneutralization to observe the changes in FSH secretion were determined in initial experiments. Pulsatile secretion of both FSH and LH was observable in the diestrous female. Two hours after inhibin immunoneutralization, the mean trough level, mean peak level, and overall mean level of FSH began to increase. The maximal increase and plateau of these parameters were observed 5 h after antiserum injection. During the period of increase, mean FSH pulse amplitude was also increased, but returned to the level observed in control (normal sheep serum-injected) animals when the parameters of trough, peak, and overall mean FSH reached their plateau levels. FSH pulse frequency was not changed at any time. These results indicate that endogenous inhibin affects only the basal parameters of FSH secretion without affecting pulsatile FSH secretion. The transient increase in FSH pulse amplitude resulted from FSH pulses being superimposed on the increasing basal FSH secretion. In contrast, immunoneutralization of endogenous inhibin rapidly increased all parameters (i.e. pulse amplitude and frequency, mean trough and peak levels, and mean plasma levels) of LH secretion. In addition, pituitary sensitivity to an exogenous LHRH challenge was increased in inhibin-immunoneutralized females in terms of stimulated LH secretion. As a result of the already increased rate of basal secretion, the actual quantity of FSH released in response to the LHRH challenge was greatly increased in the inhibin-immunoneutralized rats compared with the normal sheep serum-injected controls; however, the increase in the rate of FSH secretion stimulated by the LHRH challenge was the same in both groups. The observations from these studies collectively demonstrate that inhibin acts endogenously to suppress those parameters of gonadotropin secretion that are regulated by LHRH.