Journal: Physiological Reports

Article Title: Nephron deficit and low podocyte density increase risk of albuminuria and glomerulosclerosis in a model of diabetes

doi: 10.14814/phy2.15579

Figure Lengend Snippet: Schematic of experimental design. Female C57Bl6/J mice were fed a normal protein diet (NPD; n = 8) or a low protein diet (LPD; n = 8) from 3 weeks prior to mating and up to weaning at P21. One male and one female offspring per litter was euthanized at P21 (weaning) for assessment of glomerular and podocyte indices. Two male and two female offspring per litter were weaned onto standard laboratory chow. At 6 weeks of age, one male and one female per litter were administered i.p. injections of streptozotocin (55 mg/kg) to induce diabetes or citrate buffer (control); daily over 5 days. P, protein; F, fat; STZ, streptozotocin; CB, citrate buffer; ACR, albumin creatinine ratio; GSI, glomerulosclerotic index; BGL, blood glucose level; 6 W, 6 weeks of age; 24 W, 24 weeks of age.

Article Snippet: Creatinine was measured using the Jaffe’ reaction of alkaline picrate with creatinine (Exocell).

Techniques: Control

Journal: Physiological Reports

Article Title: Nephron deficit and low podocyte density increase risk of albuminuria and glomerulosclerosis in a model of diabetes

doi: 10.14814/phy2.15579

Figure Lengend Snippet: LPD induces growth restriction, low nephron endowment, and low podocyte endowment at weaning, but normal kidney function and histology at 24 weeks of age. (a) Pre‐weaning bodyweight (P2‐21); data analyzed by repeated measures 3‐way ANOVA for diet, sex, and time; no sex difference identified so data were consolidated and analyzed by 2‐way ANOVA for diet and time. (b) Total kidney weight (left and right kidney combined), (c) total glomerular number per kidney and (d) glomerular number per gram of bodyweight at P21; data analyzed by 2‐way ANOVA for diet and sex, no sex difference identified (e) glomerular volume, (f) podocyte number per glomerulus, and G) podocyte density at P21 and 24 weeks of age; data analyzed by three‐way ANOVA for diet, sex, and time, no sex difference identified so data were consolidated and analyzed by 2‐way ANOVA for diet and time. (h, i) Bodyweight from P21 to 24 weeks of age in male (h) and female (i) offspring; presented separately to show the identified sex difference following repeated measures 3‐way ANOVA for diet, sex, and time. (j) Left kidney weight and (k) albumin: creatinine area under the curve at 24 weeks in male and female offspring indicating the sex difference identified following 2‐way ANOVA for diet and sex. (l) glomerulosclerotic index in NPD and LPD offspring at 24 weeks of age; data analyzed by 2‐way ANOVA for diet and sex, no sex difference identified. NPD (white circles), LPD (black circles). Asterisks above data points represent significant differences between NPD and LPD; asterisks below datapoints represent significant differences between P21 and 24 weeks. In all cases, values are mean ± SD. where * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 following multiple comparisons. P, postnatal day; 24 W, 24 weeks.

Article Snippet: Creatinine was measured using the Jaffe’ reaction of alkaline picrate with creatinine (Exocell).

Techniques:

Journal: Physiological Reports

Article Title: Nephron deficit and low podocyte density increase risk of albuminuria and glomerulosclerosis in a model of diabetes

doi: 10.14814/phy2.15579

Figure Lengend Snippet: Disease susceptibility following induction of diabetes. (a) Blood glucose level presented as area under the curve (AUC) (analyzed by 3‐way ANOVA for diet, sex, and treatment) showing the significant difference between males and females. Blood glucose in male (b) and female (c) offspring across the 18‐week experimental period analyzed by 3‐way ANOVA with repeated measures for diet, treatment, and time. (d) kidney weight at 24 weeks following 18 weeks of hyperglycemia; analyzed by three‐way ANOVA for diet, sex, and treatment. Values are mean ± SD, where ** p < 0.01, *** p < 0.001, **** p < 0.0001 following multiple comparisons, differences between sexes not shown. c, control (citrate buffer); d, diabetic (STZ treatment). (e) Podocyte number per glomerulus, (f) glomerular volume, (g) podocyte density, and (h) albumin: creatinine area under the curve, in control and diabetic NPD and LPD offspring. Each dataset in e–h analyzed by three‐way ANOVA for diet, sex, and treatment. No sex difference was identified, data were consolidated and analyzed by 2‐way ANOVA for diet and treatment; in all cases values are mean ± SD, where * p < 0.05, *** p < 0.001 and **** p < 0.0001 following multiple comparisons. (i) Albumin: creatinine across the experimental period; no sex difference identified in ACR AUC, so ACR analyzed by repeated measures 3‐way ANOVA for diet, treatment, and time where * p < 0.05 between LPD control and LPD diabetic, *** p < 0.001 between LPD control and LPD diabetic, **** p < 0.0001 between LPD control and LPD diabetic, ^^^ p < 0.001 between NPD control and NPD diabetic, ^^^^ p < 0.0001 between NPD control and NPD diabetic, ## p < 0.01 between NPD diabetic and LPD diabetic offspring. (j) Glomerulosclerotic index at 24 weeks of age in control and diabetic NPD and LPD offspring, data analyzed by three‐way ANOVA for diet, sex, and treatment, no sex difference was identified, data were consolidated and analyzed by 2‐way ANOVA for diet and treatment with values presented as mean ± SD, where *** p < 0.001 and **** p < 0.0001 following multiple comparisons. (k) Linear regression of the relationship between podocyte density and glomerulosclerotic index at 24 weeks of age (18 weeks of hyperglycemia in diabetic animals). In a–k: NPD—control (white circles), LPD—control (black circles), NPD—diabetic (white squares), LPD—diabetic (black squares). C, control; d, diabetic; M, male; f, female.

Article Snippet: Creatinine was measured using the Jaffe’ reaction of alkaline picrate with creatinine (Exocell).

Techniques: Control