To clarify the functional differences among G-proteins, we investigated the localization of G(i) and G(o) in the olfactory bulb of rats by both immunohistochemical and immunochemical techniques, using purified antibodies specific to the alpha-subunits of G(i1) (G(i1)alpha), G(i2) (G(i2)alpha), and G(o) (G(o)alpha), respectively. We found that G(i2)alpha is localized exclusively in the accessory olfactory bulb, but it is present at only low levels in the main olfactory bulb. The unique pattern of immunoreactivity specific for G(i2)alpha and G(o)alpha within the glomeruli of the accessory olfactory bulb and the results of immunoassays indicate that the accessory olfactory bulb is divided into two parts: the anterior region is rich in G(i2), while the posterior region is rich in G(o). These findings suggest that the accessory olfactory bulb has two different functions. In addition, we found that the concentration of G(i2)alpha in the accessory olfactory bulb increases during puberty and reaches the adult level at 12 weeks after birth, while that in the main olfactory bulb remains constant. By contrast, the concentrations of G(o)alpha in the accessory olfactory bulb and the main olfactory bulb increase with similar kinetics. These findings suggest that G(i2) is a key protein in signal transduction in the accessory olfactory bulb, and increases in its level seem to be related to sexual maturation.