Gene regulation of cell adhesion: a key step in neural morphogenesis

A mounting body of evidence suggests that cell adhesion molecules (CAMs) play important roles in morphogenetic patterning of the nervous system. The combined factors that control the expression of CAMs during early neural development are, however, largely unknown. We have hypothesized that the coordinate expression of homeobox (Hox) and paired box (Pax) proteins in the neural axis leads to the differential expression of particular CAM genes. Following this hypothesis, we have characterized the promoters and identified cis-regulatory sequences that bind to and respond to Hox and Pax proteins in the genes for three neurally expressed CAMs - the neural cell adhesion molecule, N-CAM, the neuron-glia cell adhesion molecule, Ng-CAM, and L1. Experiments on transgenic mice carrying N-CAM promoter/lacZ reporter gene constructs indicated that mutation of either the HBS or the PBS disrupted patterning of N-CAM expression in the embryonic spinal cord. To examine the factors that restrict the expression of certain CAMs to the nervous system, we identified regulatory elements that block expression of the Ng-CAM and L1 genes in non-neural cells. We characterized a 310 base pair region of the first intron of the Ng-CAM gene containing five neural restrictive silencer elements (NRSEs) and a binding site for the Pax-3 protein. These elements silenced activity of the Ng-CAM promoter in NIH3T3 fibroblasts, but had no effect on its activity in N2A neuroblastoma cells line. Similar analyses of the L1 gene revealed a single NRSE within the second intron that was important for silencing in this cellular transfection system. To analyze the role of the NRSE in vivo, we prepared transgenic mice containing two L1 gene/lacZ constructs, one containing the NRSE and another in which the NRSE was deleted. The wild type L1lacZ transgene showed a neurally restricted pattern of expression, whereas the NRSE-mutated L1 construct showed extensive extraneural expression of the L1 gene. Thus, neural specificity of CAM expression is controlled by the NRSE. The general significance of these observations is that they connect the expression of important families of transcriptional regulators with gene products capable of direct cellular mechanochemistry.