Abstract
A photoreceptor cell line, 661W, derived from a mouse retinal tumor that expresses several markers of cone photoreceptor cells has been described earlier. However, these cells can be differentiated into neuronal cells. Here, we report that this cell line expressed certain markers specific to retinal ganglion cells such as Rbpms, Brn3b (Pou4f2), Brn3c (Pou4f3), Thy1 and γ-synuclein (Sncg), and some other markers of neuronal cells (beta-III tubulin, NeuN and MAP2). These cells also expressed Opn1mw, a cone-specific marker and nestin, a marker for neural precursor cells. Two glaucoma-associated mutants of OPTN, E50K and M98K, but not an amyotrophic lateral sclerosis-associated mutant, E478G, induced cell death selectively in 661W cells. However, in a motor neuron cell line, NSC34, E478G mutant of OPTN but not E50K and M98K induced cell death. We conclude that 661W is a retinal ganglion precursor-like cell line, which shows properties of both retinal ganglion and photoreceptor cells. We suggest that these cells could be utilized for exploring the mechanisms of cell death induction and cytoprotection relevant for glaucoma pathogenesis. RGC-5 cell line which probably arose from 661W cells showed expression of essentially the same markers of retinal ganglion cells and neuronal cells as seen in 661W cells.
Highlights
There are several types of cells in the retina organized in multiple layers which are affected by various disorders of retina that contribute to blindness worldwide
Immunostaining studies of mouse retina have revealed the expression of certain proteins in the ganglion cells, which show a strong co-relation with retrograde labeling, indicating that these proteins can be used as markers for identification of RGCs40,45,48
Immunofluorescence staining of 661W and retinal ganglion cells (RGCs)-5 cells grown on coverslips showed that Brn3b and Brn3c were expressed in all the cells (Fig. 1F and G)
Summary
There are several types of cells in the retina organized in multiple layers which are affected by various disorders of retina that contribute to blindness worldwide. The E50K mutant of OPTN is causatively associated with NTG in humans[12,35] and it has been shown to induce degeneration of RGCs in transgenic mouse models[36,37,38] Another glaucoma-associated variant of OPTN, M98K, induces cell death and Tbk1-dependent phosphorylation selectively in RGC-5 cells but not in IMR32 or HeLa cells[29,39]. These RGC-like properties of RGC-5 cells cannot be explained by the molecular marker analysis that has been done during re-characterization. We find that RGC-5 cells show very similar pattern of expression of RGC-specific and other markers that are seen in 661W cells
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