Retrograde labels by Nuclear Yellow from the female rat ventral premammillary nucleus (PMv) were most numerous in the lateral septum (LS) and the preoptic area (POA) and spread laterally into the substantia innominata. Other labels were in the diagonal band nucleus, the substantia innominata and the bed nucleus of the stria terminalis. Constant-current, single-pulse electrical stimulation of the PMv in urethan-anesthetized ovariectomized rats elicited antidromic action potentials in the cingulate cortex, in addition to the structures that contained labeled neurons. In the LS or cingulate cortex, but not in the POA, estrogen decreased antidromic activation thresholds and shortened refractory periods. The PMv is a way station that relays estrogen-regulated septal, but not preoptic, effects. The PMv also contains estrogen-sensitive fibers of passage with origins in the cingulate cortex.