Abstract
Purpose: :
Proliferative vitreoretinopathy (PVR) is a complication of retinal detachment characterized by membrane proliferation and contraction. RPE cells have been implicated as major participants in this disease. Likewise, PDGFR is found in large quantity in PVR membranes, and has been found to be intrinsic to the development of PVR in animal models. Müller cells, on the other hand, have been found to proliferate during retinal detachment and undergo de-differentiation during retinal injury. Here we examine whether un-differentiated Müller cells can express lineage markers and growth factors which could implicate them as more active participants in the pathogenesis of PVR.
Methods: :
Mouse Müller cells were isolated and cultured on 8-well cell slides coated with Type I collagen. Control cells were cultured in DMEM/F12 media with 10% FBS. Experimental cells were cultured in complete neurobasal media (rhEGF, N-2, L-glutamine, B-27) and allowed to form neurospheres. Cells were then fixed with 4% paraformaldehyde and stained with primary antibodies directed against PDGFR, PDGFRβ, RPE 65, ZO-1, GS, GFAP, cyclin D3, vimentin, ezrin, nestin, and SOX-2. The cells were then examined by conofocal microscopy.
Results: :
Control cells were found to have minimal expression of PDGFRβ, ZO-1 and GS, with strong expression of GFAP, vimentin and cyclin D3, and no expression of RPE65, ezrin, PDGFR, nestin, and SOX-2. Experimental cells had strong expression of PDGFR and β, RPE65, ZO-1, ezrin, nestin, and SOX-2, minimal expression of GS and GFAP, and no expression of vimentin.
Keywords: Muller cells • growth factors/growth factor receptors • retinal pigment epithelium