Mutation of a putative nuclear receptor binding site abolishes activity of the nestin midbrain enhancer.

Regional differences in gene expression are critical to the proper development of specialized cell types in the nervous system. The ventral midbrain is the prominent source of dopaminergic neurons, which are affected in Parkinson's disease. We have recently identified a gene regulatory element that is specifically active in ventral midbrain neuroepithelium of developing embryos. This 204-bp transcriptional enhancer is conserved within the second intron of mammalian nestin genes and contains a putative binding site for a protein of the nuclear receptor family. Our present study shows, by mutagenesis and reporter gene assay in transgenic mice, that this site is essential for enhancer function in the developing midbrain. The characterization of regulatory sites and transcription factors with specific activity in the ventral midbrain provides insight into the molecular mechanisms by which neural progenitor cells become specified towards particular neuronal differentiation pathways.