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. Author manuscript; available in PMC: 2016 Jul 19.
Published in final edited form as: Cell Metab. 2010 Oct 6;12(4):329–340. doi: 10.1016/j.cmet.2010.08.015

Figure 3. PodIRKO mice develop albuminuria and histological changes recapitulating features of diabetic nephropathy.

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A. At 8 weeks podIRKO mice become albuminuric. SDS page gel shows specific band at 66KD corresponding to Albumin only in PodIRKO mice (2μl of urine loaded for each).

B. Significant proteinuria in podIRKO mice generated on different cre promoters, raised in different environments. However there is variability both between strains and also within the groups (***p<0.001 n≥10 each group, mean +/− SEM shown).

C. Light microscopy for podocin cre podIRKO mice and controls age 8 weeks. Top panels are low power Periodic Acid Schiff (PAS) staining middle high power PAS and lower H and E staining - high power. PodIRKO (middle column) demonstrate dilated tubules containing proteinaceous casts. The glomeruli show increased mesangial matrix production. Several glomeruli appear shrunken, associated with foci of segmental glomerulosclerosis. The affected glomerular segments show adhesion to Bowman’s capsule, segmental scarring, and capillary insudative (hyalinotic) lesions. Occasional podocytes show small, hyperchromatic, condensed nuclei suggestive of apoptotic bodies (arrowed bottom middle panel). Focal glomeruli show podocyte and parietal epithelial cell swelling with large protein absorption droplets.

D. Objective histological glomerular scoring reveals significant levels of glomerulosclerosis and matrix accumulation in both the podocin and nephrin cre mice. Histological abnormality is more marked in the podocin animals. No difference in mesangial hypercellularity detected. (*p<0.05 **p<0.005 ***p<0.0005 n≥10 each group, mean +/− SEM shown). Plasma creatinine levels were measured in a subset of mice at 8 weeks. No difference was detected in the nephrin cre mice but in the podocin cre IRKO animals there was a mildly raised creatinine level in mutants when compared to controls.

E. Type IV collagen progressively accumulates in glomeruli of podIRKO mice (brown staining).

F. Podocyte apoptosis is detected using Tunel stain in 13 week podIRKO mice (nephrin promoter). Apoptotic nuclei arrowed.

G. Light microscopy of 13 week old podIRKO (nephrin promoter) showed massive amounts of matrix formation as well as increased glomerulosclerosis, mesangial expansion and protein in the tubules in comparison to the 8 week old mice. right = high power (x63 magnification), left = low power(x10 magnification) PAS staining.

H. Some 13 week old podIRKO mice had small sclerosed kidneys.

I. Transmission electron microscopy of podIRKO and control mice. PodIRKO mice have normal GFB at 3 weeks of age. By age 5 weeks the podocyte actin cytoskeleton is beginning to become deranged with broadening of foot processes (arrowed). Foot processes are completely lost by 8 weeks (arrowed). No obvious changes were detected in the GEnC.

J. Thickening of the GBM of podIRKO mice when measuring the lamina densa.

K. Multiple sections from 3 mutant mice were compared with 3 control mice. Each section had at least 10 measurements of Lamina densa thickness performed perpendicular with the edge of the GBM. **p=0.007 Mean +/− SEM shown.