Figure 1.
Glut 4 expression in the retina.
Immunoprecipitation (IP) followed by Western blotting (WB) and detection. 100 μg protein were loaded. Quantification of data is presented as the mean ± SEM of at least six animals per group. M, muscle; N, normal; 20d, 20 days diabetic rats; 45d, 45 days diabetic rats.
Figure 2.
Immunodetection of Glut 4 in the retina.
Representative photomicrographs of vertical sections of frog (A, B) and rat (C, D) retinas, showing immunoreactivity to anti Glut 4 antibody (B and D, respectively). A, C are negative controls, in which primary antibody was omitted. At least five sections of five animals were analyzed. GCL, ganglionar cell layer; INL, inner nuclear layer; ONL, outer nuclear layer; OS, outer segments; RPE, retinal pigment epithelium. Bar represents 50 μm.
Figure 3.
Glut 4 mRNA expression in the retina.
Representative sections of frog (A,B) and rat (C,D) retinas processed for in situ hybridization of Glut 4 mRNA. Low magnification photographs of alkaline phosphatase developed retinal sections showing distribution of Glut 4 mRNA (B and D, respectively). Sense probe (controls) are A and C, whereas B and D were hybridized with antisense probes. GCL, ganglionar cell layer; INL, inner nuclear layer; ONL, outer nuclear layer; OS, outer segments; RPE, retinal pigment epithelium. Bar represents 50 μm.
Figure 4.
Characterization of 14C-Glucose uptake by retinas of normal and streptozotocin-induced diabetic rats.
Tissues were incubated for 20 min in a Krebs medium with or without insulin (10 ng/ml). The effects of LY294002 (LY, 10 μM) and wortmannin (Wort, 50 nM) on incubations with added insulin are shown. Each value is the mean ± SEM of five animals. * Significantly different from control at p<0.02.
Figure 5.
Time course of insulin-induced phosphorylation of Akt.
The figure shows a Western blot of a representative experiment (top) and densitometric analyses for total (Akt) and phosphorylated-Akt (p-Akt). Retinas were incubated in Krebs medium in the absence or presence of 10 ng/ml insulin, and in the presence of LY294002 (LY, 10 μM). Data are the mean ± SEM of five different experiments, each one carried out in duplicate. * p<0.03; ** p<0.0001; + p = 0.0001; all with respect to the control (C).