We have examined the effect of electrical stimulation in the subfornical organ (SFO) on the activity of neurons in the medial septum and diagonal band of Broca (MS-DBB) which project to the median eminence in the rat. Previous studies have suggested that septal neurons with such projections are luteinizing hormone releasing hormone neurons. Extracellular single-unit recordings were obtained from 69 neurons in the MS-DBB which were antidromically identified as projecting to the median eminence. Electrical stimulation (100–400 µA) in the SFO resulted in a relatively short-latency (22.8 ± 0.8 ms), short-duration (9.2 ± 0.8 ms) activation of 85% of 41 antidromically identified MS-DBB neurons tested. One neuron was inhibited by SFO stimulation, while the remaining 12% were unaffected. Stimulation in nearby regions, including the hippocampal commissure and paraventricular nucleus of the thalamus, failed to elicit a similar excitatory response from 19 identified MS-DBB neurons. These results identify a neuronal pathway between the SFO and septal neurons which project to the median eminence, supporting a possible role for the SFO in the control of luteinizing hormone releasing hormone release and hence gonadotropin secre