This study investigated the contribution of sub-cortical afferent input to the effects of testosterone (T) on spine synapse density in the CA1 subfield of the hippocampus, in adult male rats. Gonadectomized (GDX) male rats exhibited a considerably lower density of spine synapses in the CA1 region than control, intact males. The effects of GDX were reversed by treatment with testosterone propionate (TP; 500 microg/day, for 2 days). Transection of the fimbria/fornix (FF) had no significant effect on the synaptic density in non-GDX males. However, FF transection partially inhibited the responses to TP in GDX animals. These data suggest that the effects of T on spine synapse density in the CA1 region of the male rat hippocampus are partially, but not completely, dependent on afferent sub-cortical input.