Fig. 4: Intracavernous administration of LRG1 ameliorates vascular and neurological abnormalities via LPHN2 receptor in diabetic mouse model. | Experimental & Molecular Medicine

Fig. 4: Intracavernous administration of LRG1 ameliorates vascular and neurological abnormalities via LPHN2 receptor in diabetic mouse model.

From: Latrophilin-2 is a novel receptor of LRG1 that rescues vascular and neurological abnormalities and restores diabetic erectile function

Fig. 4

a Immunostaining of PECAM-1 (green) and neuron-glial antigen 2 (NG2, red) in cavernous tissue from control and STZ-induced diabetic mice at 2 weeks after repeated intracavernous injections of Fc (negative control, 5 µg/20 µl) or LRG1-Fc (5 µg/20 µl) in Days 0 and 3 under shCon or shLPHN2 conditions (5 × 105 TU/mouse) (left and middle). Scale bars, 100 μm. PECAM-1 and NG2 (+) areas per cavernosum were quantified using ImageJ (right, n = 5). b, d, e, f and g Immunostaining of p-eNOS (b), BrdU (d), Claudin-5 (e), oxidized-low-density lipoprotein (LDL, f), TUNEL (g) (red) and PECAM-1 (green) in cavernous tissue from control and STZ-induced diabetic mice 2 weeks after repeated intracavernous injections of Fc or LRG1-Fc (5 µg/20 µl) on Days 0 and 3. Nuclei were stained with DAPI (blue, d and g). Scale bars, 100 μm (scale bars, 50 μm for only d). c Representative Western blots showing p-eNOS and eNOS in whole penile tissue from control and STZ-induced diabetic mice 2 weeks after repeated intracavernous injections of Fc (negative control, 5 µg/20 µl) or LRG1-Fc (5 µg/20 µl) on Days 0 and 3. h Quantification of p-eNOS by immunostaining and Western blotting (n = 4), the number of BrdU (+) endothelial cells per high-power field (n = 4), claudin-5 and oxidized-LDL per cavernosum (n = 4), and the number of TUNEL (+) apoptotic endothelial cells per high-power field (n = 4). i and j Immunostaining of βIII-tubulin (green) and nNOS (red) in DNB tissue from control and STZ-induced diabetic mice 2 weeks after repeated intracavernous injections of Fc (negative control, 5 µg/20 µl) or LRG1-Fc (5 µg/20 µl) on Days 0 and 3 under shCon or shLPHN2 conditions (5 × 105 TU/mouse, only for i). Nuclei were labeled with DAPI (blue). Scale bars, 25 µm for (i) and 100 μm for (j). βIII-tubulin or nNOS (+) areas in DNB tissue per high-power field were quantified using ImageJ (means ± SEM (n = 6), right). *P < 0.05; **P < 0.01; ***P < 0.001 (Student’s t test). N.S., not significant. The relative ratio of the shCon+Fc group or control group was defined as 1.

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