The vomeronasal system projects to the accessory olfactory bulb (AOB), to the medial (Me) and posterior medial cortical nuclei (PMCN) of the amygdala, to the bed nucleus of the stria terminalis (BNST), and to other central structures shown to be important in mating behavior, including the medial preoptic area (MPOA). In these experiments c-fos expression was used as a marker of neural activity to identify the contribution of vomeronasal sensory input during mating behavior in male golden hamsters, either intact or with vomeronasal organs removed (VNX). Inexperienced hamsters were either stimulated with a receptive female and allowed to mate, exposed to female hamster vaginal fluid (HVF), which contains stimuli known to act through the VN system, or placed in a clean cage alone. Densely stained Fos-positive nuclei were evident in mated animals in the central VN pathway [AOB, Me, posterior medial BNST (pmBNST)] and a VN target area (MPOA). HVF-exposed animals showed Fos expression in the AOB, Me, and BNST but not MPOA. Unstimulated animals showed almost no activation. Most VNX animals exposed to females did not mate, but performed intense chemoinvestigation. They had few Fos-positive nuclei in any of these areas except the caudal pmBNST. A few VNX animals that did mate had patterns of Fos activation that were similar but less intense than those of intact mating animals, suggesting a selective activation of VN central pathways during mating regardless of VN sensory input. The main olfactory system showed low levels of Fos expression in all animals (stimulated and unstimulated). Fos expression in the MPOA and rostral pmBNST was seen only in mated animals, suggesting that these regions are concerned with mating performance or its consequences, rather than the chemosensory input that triggers it. Fos expression in the caudal encapsulated pmBNST was evident in all groups of animals that performed chemosensory investigation, regardless of VN status or mating, suggesting that this region either directs or responds to chemosensory investigation.