Neurog2 regulates Isl1 to modulate horizontal cell number

The population sizes of different retinal cell types vary between different strains of mice, and that variation can be mapped to genomic loci in order to identify its polygenic origin. In some cases, controlling genes act independently, whereas in other instances, they exhibit epistasis. Here we identify an epistatic interaction revealed through the mapping of quantitative trait loci from a panel of recombinant inbred strains of mice. The population of retinal horizontal cells exhibits a two-fold variation in number, mapping to quantitative trait loci on Chromosomes 3 and 13, where these loci are shown to interact epistatically. We identify a prospective genetic interaction underlying this, mediated by the bHLH transcription factor Neurog2, at the Chromosome 3 locus, functioning to repress the LIM homeodomain transcription factor Isl1, at the Chromosome 13 locus. Using single and double conditional knockout mice, we confirm the countervailing actions of each gene, and validate in vitro a critical role for two single nucleotide polymorphisms in the 5′UTR of Isl1, one of which yields a novel E-box, mediating the repressive action of Neurog2.