Kidney phenotype at 30 weeks of age.

<p>Values are means ± SEM; n, number of mice;</p>*<p>P<0.05 between diabetic and control animals.</p

Gene expression in cardiac tissue.

<p>Cardiac gene expression at 10 weeks of age; mRNA levels in diabetic mice relative to controls (assigned as 1.0) normalized to β-actin mRNA level. Values are means ± SE; n, number of mice; TGFβ, transforming growth factorβ; MCP-1, monocyte chemoatactic protein-1; IL1β, interleukin 1β; TNFα, tumor necrosis factor-α; CTGF, connective tissue growth factor.</p>*<p>P = 0.001 between diabetic and control animals.</p

Fasting blood glucose levels and urine output in male Akita mice.

<p>(A) FVB/NJ-<i>Ins2</i>^+/C96Y mice developed sustained hyperglycemia by 4 weeks of age, which persisted throughout the duration of the study. (B) Urine output in FVB/NJ-<i>Ins2</i>^+/C96Y was significantly increased by 8 weeks of age and further increased during the study period. Black bars = control (n = 6 to 8); white bars = Akita (n = 6 to 8). Data are mean ± SEM. *P<0.05 †P<0.001 or **P<0.01 vs. age-matched controls.</p

Cardiac injury in male Akita mice.

<p>(A–B) Panels A and B show representative photomicrograph of heart sections from controls (A) and diabetic mice (B) at 10 weeks of age. Examination of Akita mice hearts revealed mild interstitial fibrosis (green) and focal inflammatory infiltrates (arrows). Tissue sections were stained with Masson trichrome and the magnification was 400×. (C) Expression of mRNA for β-myosin heavy chain (β-MHC) was enhanced in 10-week old Akita mice. Levels of mRNA were expressed relative to controls (assigned as 1.0) and normalized to β-actin mRNA level. Black bars = control (n = 5); white bars = Akita (n = 5). Data are mean ± SEM. *P<0.05 vs. age-matched controls.</p

GBM and proximal tubule basement membrane thickness at 30 weeks of age.

<p>Values are means ± SEM; n, number of mice.</p

Podocyte injury in normoalbuminuric diabetic Akita mice.

<p>(A) Albuminuria was not significantly different in Akita mice and controls at 4 weeks of age. (B) In contrast, urinary nephrin excretion was increased in normoalbuminuric 4-week old Akita mice compared to control animals. (C) There was a significant increase in podocyte apoptosis in Akita mice compared to age-matched controls without a change in podocyte number. Black bars = control (n = 6 to 8); white bars = Akita (n = 6 to 8). Data are mean ± SEM. *P<0.05 or †P<0.001 vs. age-matched controls.</p

Kidney weight and glomerular filtration rate (GFR) in male Akita mice.

<p>(A) Kidney-to-body weight ratio was significantly increased in diabetic Akita mice (n = 6 to 8) compared to controls (n = 6 to 8) at 12 and 20 weeks of age. (B) There was a significant increase in GFR at 12 weeks of age in diabetic Akita mice (n = 3 to 5) compared to controls (n = 5). Black bars = control; white bars = Akita. Data are mean ± SEM. *P<0.05 or **P<0.001 vs. age-matched controls.</p

Urinary albumin excretion (UAE) in male Akita mice.

<p>(A) UAE was significantly elevated compared with controls in diabetic Akita mice by 8 weeks of age. By 20 weeks of age, there was a >10-fold increase in albuminuria in diabetic mice compared to control animals. (B) Albumin-to-creatinine ratio (ACR) was also significantly increased in diabetic mice compared to age-matched controls. (C) There was a linear relationship between 24-hour UAE and ACR in all mice (R² = 0.71; P<0.0001). Black bars = control (n = 6 to 8); white bars = Akita (n = 6 to 8). Black circles = control; white circles = Akita. Data are mean ± SEM. *P<0.05 or †P<0.0001 vs. age-matched controls.</p

Body weight.

<p>Values are means ± SEM; n, number of mice.</p

Podocyte injury in albuminuric diabetic Akita mice.

<p>(A) Urinary nephrin excretion was significantly enhanced in Akita mice compared to controls at 16 and 20 weeks of age. (B) There was a significant linear relationship between urinary nephrin excretion and albuminuria in all mice (R² = 0.57; P<0.0001). (C) Podocyte number was significantly decreased in Akita mice at the 20-week time point. In contrast, podocyte apoptosis was difficult to detect in either Akita mice or control animals. Black bars = control (n = 6 to 8); white bars = Akita (n = 6 to 8). Black circles = control; white circles = Akita. Data are mean ± SEM. *P<0.05 or †P<0.01 vs. age-matched controls.</p