Abstract
Epidermal growth factor (EGF) signaling has been implicated in the regulation of the differentiation and proliferation of retinal progenitors. We assessed how different levels of EGF signaling, achieved either by increasing receptor expression or via addition of the exogenous ligand, or an increase in both, can affect the differentiation of progenitors in the first week of postnatal retinal development in the model system of retinal explants (REs). Proliferating progenitor cells in REs were infected with either the control CLV3/ESR-related peptide family (CLE)-green fluorescent protein (GFP)- or with EGF receptor (EGFR)-GFP-expressing retrovirus, and grown in the control medium or in the presence of exogenous EGF (10 ng/mL). The differentiation of infected cells into Muller glia (Sox9+), rod photoreceptors (rhodopsin+) and horizontal cells (calbindin+) was analyzed. In all the examined conditions, infected cells differentiated into Muller glia and rod photoreceptors that normally develop postnatally. Horizontal cells finished their development during the embryonic stages and progenitors infected with control-GFP virus did not differentiate into GFP+/calbindin- in either control or EGFsupplemented medium, however, cells infected with EGFR-GFP differentiated into horizontal cells (GFP+/calbindin+) in both culture conditions. These results imply that altering the levels of EGFR and/or the amount of the EGF ligand can overcome progenitor competence restriction.
Highlights
The vertebrate neural retina is comprised of six types of neurons: rod and cone photoreceptors, amacrine cells, retinal ganglion cells (RGCs), horizontal cells, bipolar cells and one type of glia, Müller glia
Horizontal cells finished their development during the embryonic stages and progenitors infected with control-green fluorescent protein (GFP) virus did not differentiate into GFP+/calbindin- in either control or EGFsupplemented medium, cells infected with EGF receptor (EGFR)-GFP differentiated into horizontal cells (GFP+/calbindin+) in both culture conditions
P0 progenitors infected with control-GFP and EGFR-GFP retrovirus differentiate into Müller glia and rod photoreaffects cell fate choice of P0 postna- ceptors in either control or Epidermal growth factor (EGF)-supplemented medium
Summary
The vertebrate neural retina is comprised of six types of neurons: rod and cone photoreceptors, amacrine cells, retinal ganglion cells (RGCs), horizontal cells, bipolar cells and one type of glia, Müller glia. To understand how different levels of EGF signaling affect the development of specific cell types in postnatal retina, we cultured P0 REs under four different conditions: (i) progenitors infected with control-GFP and grown in the control media; (ii) progenitors infected with control-GFP and grown in the presence of the exogenous EGF (10 ng/mL); (iii) progenitors infected with EGFR-GFP and grown in the control medium; (iv) progenitors infected with EGFR-GFP and grown in the presence of exogenous EGF (10 ng/mL) (the schematic overview of the experimental paradigm is presented in Supplementary Fig. S1A).
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