NEUROTROPHIN-4/5 PROMOTES SURVIVAL AND DIFFERENTIATION OF RAT STRIATAL NEURONS DEVELOPING IN CULTURE

Cultures of dissociated striatal neurons from fetal rats were prepared, and were grown in the presence of neurotrophin-4/5 (NT-4/5) as well as the other known neurotrophins, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3). We found that acute administration of NT-4/5 to 7-day-old cultures stimulates the hydrolysis of phosphatidylinositol, an event involved in neurotrophin signal transduction. Growth of striatal cultures in the presence of NT-4/5 resulted in increased cell survival, as indicated by elevations in cell number, protein content, and a measure of mitochondrial enzyme activity (MTT assay). NT-4/5 increased GABA uptake and staining intensity in these cultures, as indicated by GABA immunocytochemistry, indicating a trophic action on GABAergic neurons, the predominant neuron type in the striatum. To further identify responsive cell populations we analysed for calretinin, a calcium-binding protein known to colocalize with GABA in a number of neuronal cells. In cultures prepared from rats of embryonic day 15, NT-4/5 strongly increased the number of calretinin-positive cells as well as calretinin levels, as determined by Western blot analysis. When the cultures were prepared from embryonic day 18 rats, NT-4/5 very strongly increased the morphological differentiation of calretinin-positive cells, whereas the increase in cell number was less prominent. All effects produced by NT-4/5 were mimicked by BDNF with similar potency. NT-3 was less effective than NT-4/5 and BDNF, and its effects were limited to cultures prepared from embryonic day 15 rats, suggesting a role in the regulation of cell survival at early developmental stages. NGF did not affect any of the measured parameters. Our findings identify NT-4/5 as potent neurotrophic factor for striatal neurons, able to promote their survival and differentiation.