The epigenetic manipulation of precursors may provide data to elucidate the potential interactions among these cells in different brain regions. However, the response to epigenetic signals is modulated by the environment in which the cells are manipulated. Therefore, data regarding the action of a particular factor must be considered in the light of a specific system. To compare septal and striatal precursors, we have tested the effect of nerve growth factor (NGF) on the proliferation and neuronal differentiation of epidermal growth factor (EGF)-responsive cells from these brain regions. Precursors were cultivated as 'neuropheres' in serum free medium (SFM) to which NGF was added. NGF did not support the proliferation of EGF-generated precursors so that no differences in the cell magnitude with respect to control cultures were observed. Differentiation of precursors in SFM plus 1% fetal bovine serum (FBS) on poly-D-lysine showed that the neuron number was increased two-fold in septal cultures treated with NGF but not in those from striatum. A quantitative evaluation of the soma surface and the number of primary neurites showed differences between both populations of precursor-generated neurons. In addition, we also observed no influence of NGF on these parameters of cellular morphology. Thus, taken together these results seem to indicate that at this developmental stage in which these populations of precursors were isolated, heterogeneities exist between them, which is probably related to their origin and/or functional roles in vivo.

ResumenEn este trabajo se describe la acción del factor de crecimiento epidérmico (FCE) sobre las células del estriado embrionario en un sistema de cultivo disociado de neuronas y glías. El cultivo de células se preparó a partir de embriones de ratas de 16-17 días. En el sistema de cultivo empleado, la población celular fue cultivada durante 20-24 horas en un medio que contenía suero y, posteriormente, fue tratada con 20 ng/mL del FCE en un medio libre de suero. La eliminación del suero en este período inicial de desarrollo provocó una disminución apreciable de las células vivas en los cultivos tratados y en los controles. Al parecer, la población de células sobrevivientes estaba integrada, en su mayoría, por precursores celulares teniendo en cuenta su capacidad proliferativa. La acción del FCE sobre las células se manifestó en un aumento del número de células debido fundamentalmente a un estímulo de la proliferación de los precursores neuronales y astrocitos. Este efecto estuvo acompañado por una reducción de la diferenciación morfológica neuronal cuando se comparó con los cultivos controles. En los cultivos, a los 16 días, la detección de la actividad específica de la colina acetiltrasferasa evidenció la diferenciación de una subpoblación neuronal colinérgica, las cuales respondieron al tratamiento con el factor de crecimiento nervioso con un aumento de la actividad de la enzima. SummaryThis paper describes the effect of epidermal growth factor (EGF) on embryonic striatal cells in mixed neuronal-glial cultures prepared from 16-17 day old rat embryos. In the culture system employed, cell population was cultured for 20-24 hours in serum supplemented medium prior to being treated with 20 ng/mL epidermal growth factor (EGF) serum-free medium. This early withdrawal of serum from the culture medium caused an appreciable decline of viable cells in both control and treated cultures. It seems that the majority of surviving cells were precursors, taking into account their proliferative capacity. EGF action on cell population provoked an increase in cell numbers, mainly due to the stimulation of neuronal precursor and and astrocyte proliferation. This increase in cell proliferation was accompanied by neuronal morphological differentiation delay when compared to control cultures. Choline acetyltransferase specific activity detected in 16-day old cultures, showed the differentiation of a cholinergic neuronal subpopulation, which responded to nerve growth factor treatment with enhanced enzyme activity.

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