Angiotensin II (AII) immunoreactive cells and fibers were identified in the subfornical organ (SFO) of the rat. Cells were distributed in an annulus around the periphery of the SFO and were most visible in the Brattleboro rat treated with colchicine. Fibers were observed in a plexus, located centrally within the ring of cells, and knife-cuts suggested that they arise from parent cell bodies lying outside of the SFO. Studies combining immunohistochemistry with retrograde transport identified the perifornical zone of the lateral hypothalamus, the rostral zona incerta, and the nucleus reuniens of the thalamus as the source of AII-stained inputs to the SFO, and the region of the median preoptic nucleus as a recipient of AII-immunoreactive SFO efferents. It is suggested that these biochemically defined connections of the SFO participate in the central neural control of fluid balance.