Distribution and developmental expression of the nerve terminal protein NT75 in the rat cerebellum

Previous studies of the nerve terminal protein NT75 in the developing spinal cord have suggested an association between the appearance of NT75 immunoreactivity and the process of synaptogenesis. To examine the time course of NT75 expression further, the current study compared the localization of NT75 and the synaptic vesicle protein synaptophysin in the adult and developing rat cerebellum and in cerebellar tissue cultures. In the adult cerebellum, dense NT75 staining is confined to the molecular layer, where it is associated with parallel fiber endings of cerebellar granule cells. During development, NT75 immunoreactivity is first detectable in the cerebellar cortex as a dense band of staining in the deepest portion of the molecular layer at postnatal day 10. The stained zone expands to occupy a progressively greater portion of the molecular layer until about postnatal day 20. Synaptophysin staining occurs in granule cell processes earlier than NT75 and is found throughout the molecular layer by postnatal day 7. Quantitatively, rapid increases in both NT75 and synaptophysin occur in the first three postnatal weeks, with NT75 activity reaching levels exceeding the adult value by 50% over postnatal days 20 through 30, whereas synaptophysin plateaus at near adult levels by postnatal day 20. In cerebellar cultures, NT75 staining in neurites develops over several days, increasing coincidentally with development of synaptic contacts, whereas synaptophysin staining is already present in most neurites after only 1 day in vitro. The results indicate that NT75 expression in developing cerebellar granule cell nerve terminals is closely associated with the appearance of mature nerve terminals, suggesting that the protein may have a role in the formation/stabilization of the synaptic ending or in the mechanisms of synaptic transmission.