Differential expression of G proteins (Gi alpha 2 and G(o alpha) and the separate central projections of Gi alpha 2- and G(o alpha)-immunoreactive (ir) vomeronasal receptor neurons were investigated in the mouse and rat using immunocytochemical methods. In the vomeronasal organ (VNO), receptor neurons with their cell bodies located in the middle layer (middle 1/3) of the vomeronasal sensory epithelium express Gi alpha 2. Axons of these Gi alpha 2-ir neurons can be followed from VNO to the anterior part, but not the posterior part, of the nerve-glomerular (N-GL) layer of the accessory olfactory bulb (AOB). Another population of receptor neurons, which are located in the deep layer (basal 1/3) of the vomeronasal sensory epithelium, express G(o alpha), and axons of the G(o alpha)-ir neurons can be traced to the posterior part, but not the anterior part, of the N-GL layers of the AOB. The axons of the two subclasses of receptor neurons are intermingled near the VNO and become segregated as they enter the AOB. Removal of the AOB results in retrograde degeneration of both Gi alpha 2-ir and G(o alpha)-ir receptor neurons in the VNO. These results suggest that at least two subclasses of receptor neurons exist in the VNO: the Gi alpha 2-ir neurons in the middle layer and the G(o alpha)-ir neurons in the deep layer of the VNO. The Gi alpha 2-ir neurons in the middle layer of the VNO project to the anterior part of the AOB, while the G(o alpha)-ir neurons in the deep layer of the VNO project to the posterior half of the AOB. These results are similar to our previous observations in the gray short-tailed opossum, suggesting that the existence of at least two subclasses of receptor neurons in the vomeronasal epithelium with differential projections to the AOB is a conserved feature among mammals.