Neurotrophins and Basic Fibroblast Growth Factor Induce the Differentiation of Calbindin-Containing Neurons in the Cerebral Cortex

Abstract

Lineage studies have recently shown that the expression of calcium-binding proteins in neurons of the cerebral cortex is not genetically programmed and is likely to be induced by external factors. Current hypotheses suggest that basic fibroblast growth factor (bFGF) and a number of neurotrophins play important roles in the proliferation and differentiation of cortical progenitor cells to a particular lineage. Using a dissociated cell culture system, we found that bFGF and the neurotrophins brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), and nerve growth factor differentially affect the expression of the calcium-binding protein calbindin in selective neuronal subpopulations in the developing cerebral cortex. Specifically, BDNF and NT-3 greatly promoted the morphological differentiation of a relatively small, early-generated population of GABAergic neurons and induced the expression of calbindin in these cells. Furthermore, treatment with BDNF, NT-3, and bFGF produced an two- to threefold increase in the number of newly generated calbindin-positive neurons. The effect of bFGF was more striking in earlier (E14) than later (E16) ages, whereas the action of neurotrophins was independent of the age from which the cultures were prepared. Switching experiments combined with BrdU incorporation have suggested that NT-3 acts on postmitotic neurons rather than on proliferating progenitors to induce calbindin expression and that its action is mediated via trk receptors. Application of retroviral vectors in culture resulted in the presence of neuronal clones that were predominantly heterogeneous with regard to calbindin expression, suggesting, in agreement with our earlierin vivostudies, that the expression of this calcium-binding protein is not lineage dependent. Our results characterize the roles of BDNF, NT-3, and bFGF in the expression of calbindin in developing neocortical neurons.