Induction of the c-fos protein product (Fos) was used to immunocytochemically identify oxytocin (OT) neurons that may be activated during copulatory interactions. Fos induction w a s quantified in sexually-experienced male rats after either (a) exposure to a testing arena recently vacated by an estrous female, (b) copulatory interactions such as mounting and intromission without ejaculation, or (c) mounting and intromissions culminating in ejaculation. In the parvocellular regions of the paraventricular nucleus of the hypothalamus (PVN), t h e number of neurons expressing Fos increased following either intromission (53%) or ejaculation (124%). Significant, but less striking, increases in the number of cells expressing Fos were noted in magnocellular regions of the PVN where intromission resulted in a 13% increase and ejaculation in a 49% increase in Fos. The number of perikarya immunoreactive for OT and AVP did not differ as a function of increasing sexual contacts. In control (novel arena) males, 33-73% of the Fos labeling occurred in OT cells. Sexual interactions did not enhance the number of double-labeled cells in most parvocellular regions. However, in lateral parvocellular regions located in the most caudal aspects of the PVN, 31% of the Fos-positive cells occurred in OT neurons in ejaculated males, while in control males none of the OT cells were double-labeled. This PVN subdivision is known to consist of neurons that project to the brain stem and spinal cord at lumbar levels which contain motor neurons that regulate penile reflexes. The present data suggest a possible neurochemical circuit which incorporates oxytocinergic neurons in the mediation of masculine sexual responses Several studies have indicated that in addition to pituitary release from magnocellular axon terminals, oxytocin (OT) from parvocellular neurons of the paraventricular nucleus of the hypothalamus (PVN) can be released within the central nervous system (1-3). This intra-cerebral network of oxytocinergic projections may be involved in the regulation of male sexual behavior (4-9), and in particular penile erection (10-13). Although both parvo-and magnocellular OT cells have been shown to be trans-synaptically stimulated by multiple neuronal circuits during female reproduction and lactation (14, 15), little is known about the specific stimulus-coupled events affecting OT cells (in the PVN) in the male brain.The discovery of immediate early genes (IEG) in the central nervous system has made possible the cellular study of complex stimulus-coupled events and the activation of gene expression (1 6). The protein product of the c-fos IEG is rapidly, although transiently. induced and has been implicated in the control of genomic events (acting as a transcription factor) leading to prolonged functional changes in specific neurons in response to various external stimuli (17). Thus, Fos immunocytochemistry is a useful method for the neurochemical mapping of circuits involved in signal transduction and gene expression in specific neurona...