Expression of inherent neuronal shape characteristics after transient sensitivity to epigenetic factors

Abstract

We investigated effects of different substrates and culture media on the early morphological differentiation of rat neocortical neurons in culture. In particular, we examined the effects of homotypic astrocytes, the adhesive glycoprotein laminin and the polycationic substrate poly- l-lysine, as well as diffusible astrocyte-derived conditioned medium factors and serum on (1) soma area, (2) total neuron area and (3) primary neurite number. To assess variations in morphological reactions of neurons with a defined neurotransmitter phenotype, we analyzed the differentiation of GABAergic neurons. The morphology of young neocortical neurons was dramatically affected by both substrate and culture medium. Replacement of the astrocytic monolayer or the astrocyte-conditioned medium by other substrates or non-conditioned medium, respectively, was accompanied by (1) spreading and flattening of neuronal somata, (2) a marked decrease in total neuron area and (3) an increase in the number of primary neurites. The various morphological parameters studied exhibited different sensitivities to changes of these external factors. Moreover, the influences of epigenetic factors on the generation of primary neurites depended on the transmitter phenotype of the neuron. The induced morphological alterations were transient. At the end of the first week in culture, the surviving neurons underwent substantial remodeling of their morphology leading to an expression of in vivo shape characteristics. These observations suggest that despite an early, transient sensitivity to environmental influences, the neuronal differentiation with respect to the morphological parameters studied in culture is to a large degree determined by intrinsic factors.