Figure 4. Diabetes and overexpression of proNGF induced expression of p75^NTR in vivo and in vitro.

A. WB analysis showing 1.9-fold increase in the expression of p75^NTR in diabetic rats as compared with the controls (n = 4–6). B. WB analysis of rat retinal lysate showed significant increase in p75^NTR expression in rats electroporated with proNGF as compared with those electroporated with GFP (n = 4). C. Representative images of rat retina sections showing prominent immunolocalization of p75^NTR in GCL and INL in proNGF overexpressing retinas as compared with GFP-controls (400× magnification). D. Representative images of rat retina sections showing colocalization between p75^NTR in the ganglion cell layer (green) and the specific neuronal marker Thy-1 (red) in the upper pannel or with the specific RGC marker Brn-3a (red)in the lower pannel (400× magnification). E. Western blot analysis shwoing 1.6-fold increase in the expression of p75^NTR in RGC-5 cells treated with proNGF as compared with the controls (n = 4). F. Real-time PCR analysis showing that proNGF induced p75^NTR mRNA expression in freshly isolated primary RGC as compared with the control group. Samples of primary RGC cultures were pooled from 4-different cultures. Treatment with Y27632 significantly reduced p75^NTR expression in vivo and in vitro. G. Representative images showing colocalization (yellow arrow heads) of RGC that expressed p75^NTR (red) and the specific RGC marker Brn3a (green) in isolated mixed neuronal cultures from adult mice. * = significant difference as compared with the rest of the groups at p<0.05. C, control; GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; ONL, outer nuclear layer. C, control.