Figure 8.

Induction of PS expression in glomeruli attenuates podocyte injury and albuminuria in OVE26 mice. Four mice received injection of GFP-expressing rAAV9 into the left kidney for 3 weeks while the right kidneys were used as controls. (A) Representative images of immunofluorescence costaining between eGFP and synaptopodin are shown to compare the eGFP expression and localization between right kidney (control) and left kidney (rAAV9-eGFP injected). (B) Quantification of fluorescence intensity of eGFP/synaptopodin ratio between right and left kidneys from these mice (n=4; ***P<0.001 compared with the right kidneys). (C) Diabetic OVE26 mice at age 10 weeks were injected with the rAAV9-PS in the left kidneys and were euthanized 5 weeks postinjection. Representative image of immunostaining for PS and synaptopodin shows extensive overlap. (D) Pros1 mRNA levels were compared between left kidney (rAAV9-PS) and right kidney (control) by real-time PCR (n=4; **P<0.01 compared with right kidneys). (E) Representative images of PAS-stained kidneys. (F) Quantification of mesangial fraction (n=4; *P<0.05 compared with right kidneys). (G and H) Representative image of (G) WT-1 immunostaining and (H) quantification are shown (n=4; **P<0.01 and ***P<0.001 when compared between indicated groups). (I and J) In another set of experiments, OVE26 mice at age 10 weeks received either rAAV9-PS or rAAV9-eGFP in both kidneys. (I) Real-time PCR analysis confirmed an increase of PROS1 expression in the kidneys of mice injected with rAAV9-PS as compared with those injected with rAAV9-eGFP. (J) Urinary albumin-to-creatinine ratio (UACR) was determined weekly in these mice for 5 weeks (n=4; ***P<0.001 compared with between indicated groups). AU, arbitrary units; DAPI, 4′,6-diamidino-2-phenylindole; eGFP, enhanced green fluorescent protein; OVE-Control, OVE26 injected with rAAV9-eGFP; OVE-PS, OVE26 mice injected with rAAV-PS; WT-Control, wildtype injected with rAAV-eGFP; WT-PS, wildtype injected with rAAV9-PS.