Arginine-vasopressin (AVP) has been implicated as a putative central neurotransmitter or neuromodulator in some brain functions. This study demonstrates binding of [3H]AVP to rat brain homogenates that is pH and temperature dependent, is saturable (Kd = 0.77 nM, Bmax = 0.374 pmol/mg) and reversible. A number of AVP analogues competitively displaced the [3H]AVP binding, indicating that central AVP binding sites may have a resemblance to the peripheral (V1) AVP vasopressor receptor. Homogenate binding occurred predominantly in the microsomal fraction (P3) of the hypothalamus while in the hippocampus and septum binding was predominantly in the synaptosomal fraction (P2). Autoradiographic methods showed displaceable [3H]AVP binding in the lateral septum, amygdala, supraoptic, paraventricular and suprachiasmatic nuclei of the hypothalamus supporting the results of homogenate binding in preparations of these regions.