For correct functioning of the nervous system, the appropriate number and complement of neuronal cell types must be produced during development. However, the molecular mechanisms that regulate the production of individual classes of neurons are poorly understood. In this study, we have investigated the function of the thrombospondin-1-like glycoprotein, Nel (Neural epidermal growth factor-like), in the generation of retinal ganglion cells (RGCs) in chick. During eye development, Nel is strongly expressed in the presumptive retinal pigment epithelium and RGCs. Nel overexpression in the developing retina by in ovo electroporation has increased the number of RGCs, whereas the number of displaced amacrine cells has been decreased. Conversely, knockdown of Nel expression by transposon-mediated introduction of RNAi constructs has resulted in a decrease in the RGC number and an increase in the number of displaced amacrine cells. Modifications of Nel expression levels do not appear to affect proliferation of retinal progenitor cells, but they significantly alter the progression rate of RGC differentiation from the central retina to the periphery. Furthermore, Nel protects RGCs from apoptosis during retinal development. These results indicate that Nel positively regulates RGC production by promoting their differentiation and survival during development.