In vitro analysis of a mammalian retinal progenitor that gives rise to neurons and glia.

In vivo lineage studies have shown that retinal cells arise from multipotential progenitors whose fates are regulated by cell-cell interactions. To understand the mechanism underlying their maintenance and differentiation, we have analyzed the differentiation potential of progenitors derived from embryonic rat retina in vitro. These progenitors proliferate and remain undifferentiated in vitro in the presence of epidermal growth factor (EGF) and display properties similar to stem cells. In addition to expressing nestin, the neuroectodermal stem cell marker, retinal progenitors are multipotential. Upon withdrawal of EGF and addition of serum, the progenitors downregulate the expression of nestin and express cell-type specific markers corresponding to neurons and glia. In addition to expressing cell-type specific markers, retinal progenitors and their progeny could be distinguished on the basis of their distinct voltage gated current profile. A proportion of progenitors is lineage restricted and the fate of these cells can be influenced by the microenvironment, suggesting that stage-specific interactions mediated by the local environment influence the progression of progenitors towards acquisition of differentiated phenotypes.

Involvement of Mash1 in EGF-mediated regulation of differentiation in the vertebrate retina.

It is believed that signaling through the epidermal growth factor (EGF) receptor plays a critical role in the development of Drosophila eyes. In the present study we have analyzed the role that EGF-mediated signaling plays in vertebrate retinal development. We have observed that during late retinal neurogenesis EGF delays rod photoreceptor differentiation and that this effect of EGF involves the modulation of expression of a homologue of Drosophila proneural genes, Mash1. EGF causes a significant decrease in Mash1 expression and an increase in the proportion of proliferating cells in the retina in vitro. The decrease in Mash1 expression is accompanied by a concomitant decrease in opsin expression, a marker for overt rod photoreceptor differentiation. Withdrawal of EGF leads to an increase in both Mash1 and opsin expression; however, the onset of expression of Mash1 precedes that of opsin. Our study identifies a proliferative intermediate precursor, characterized by Mash1 expression, that is the target of EGF-mediated suppression of rod photoreceptor differentiation. Based on the evolutionarily conserved roles of EGF- and Notch-mediated signaling in the delay of differentiation in proliferating precursors we propose that these distinct signaling mechanisms act in concert to ensure the fidelity of the strict temporal and spatial nature of cell fate determination in the retina.