Abstract: Levels of neurotrophin‐3 markedly decrease in the rat cerebellum after the first 10 days of life, suggesting an importance during early development. To further examine the effect of neurotrophin‐3 on the developing cerebellum, we injected a monoclonal antibody against neurotrophin‐3 into the lateral ventricle of 7.5‐day‐old rats. The resultant depletion of neurotrophin‐3 caused a significant decrease in cerebellar wet weights noted at 7 and 23 days thereafter. Other changes noted 48 h after injection of monoclonal antibodies against neurotrophin‐3 included reduced incorporation of bromodeoxyuridine into granule neurons in the external germinal layer, an elevated density of atrophic neurons that had just migrated under the Purkinje cell layer, and an increased number of apoptotic neurons in the internal granule cell layer. These changes were limited to the central lobe. The concentration of neurotrophin‐3 protein in the posterior region, including the central lobe, was about four‐ and threefold higher than that in the anterior region of the cerebellum of 9.5‐ and 30‐day‐old rats, respectively. Immunocytochemical examination showed higher amounts of neurotrophin‐3 protein in the central lobe than in the anterior lobe. Our results provide evidence that neurotrophin‐3 regulates the proliferation of granule precursors and supports the survival of mature granule neurons in restricted lobules, suggesting an involvement in limited regions at a specific stage in development of the rat cerebellum.