In most murine models of renal disease and human renal disease, testosterone worsens renal disease severity, thus protection against renal disease by high testosterone in lupus mice, would be contradictory to almost all other models of renal disease

In most murine models of renal disease and human renal disease, testosterone worsens renal disease severity, thus protection against renal disease by high testosterone in lupus mice, would be contradictory to almost all other models of renal disease. analysis was used to compare trends in animal survival. Results Effect of ER deficiency on albuminuria in NZM2410 mice NZM2410 mice of various ER genotypes were placed in metabolic cages and 24-hour urine samples were collected. Urine was analyzed by ELISA for albumin as a measure of renal disease. Wildtype female NZM2410 mice, as expected (Fig. 1A), developed severe albuminuria ( 1 mg/mouse/day) by 26 weeks of age, while ER heterozygote littermate females developed severe albuminuria at 30 CTNND1 weeks of age. ER homozygote knockout NZM2410 females displayed significant protection from development of albuminuria with 24-hour albumin excretion levels never exceeding 100 g/mouse/day out to 32 weeks of age. There was a statistically significant decrease in albuminuria in ER knockout female NZM2410 mice compared to both heterozygote and wildtype ER NZM2410 knockout mice. A significant decrease in albuminuria was also observed comparing female heterozygote to wildtype mice indicating an ER gene dosing effect on albuminuria in NZM2410 female mice. ER deficiency resulted in a trend towards decreased albuminuria in male NZM2410 mice that did not reach statistical significance (Fig. 1B). Open in a separate window Figure 1 Proteinuria in ER deficient NZM2410 lupus mice. NZM females with differing ER genotype were compared for excretion of urine albumin at the various ages indicated. (A) ANOVA analysis demonstrated a significant decrease in proteinuria in ER deficient females compared to wildtype and heterozygote mice =0.041) and heterozygote ER female mice (= 0.032) as shown in Figure 2A. There was no significant difference in renal pathology scores between heterozygote and wildtype mice. This lack of a difference may reflect that the most severely affected wildtype mice had died prior to 32 weeks, thus falsely lowering the pathology scores in this group. ER knockout mice showed a decrease in all renal pathology parameters scored indicating a global impact on renal disease (Table 1). In male NZM2410 mice, ER homozygote and heterozygote knockout mice had trends toward lower renal pathology scores than wildtypes, but did not reach a significant difference (=0.08 and 0.10 respectively) when compared to wildtype (Fig. 2B). Open in a separate window Figure 2 Renal pathology in ER deficient NZM2410 lupus mice. Total glomerular pathology scores were determined for female NZM mice of various ER genotypes. (A) ER null females had significantly lower pathology score compared to both wildtype and ER heterozygotes. em p /em 0.05 by MannCWhitney analysis. (B) Male littermates of various ER status were also compared for glomerular scores with no significant difference. (C) ER null females had a trend towards lower glomerular scores but did not reach statistical significance. Littermate male NZM mice had no difference in pathology regardless of ER genotype (D). Table 1 Percentage of female NZM mice with renal pathologic parameters thead th valign=”bottom” align=”left” rowspan=”1″ colspan=”1″ ER /th th valign=”bottom” align=”left” rowspan=”1″ colspan=”1″ HC (3+) /th th valign=”bottom” align=”left” rowspan=”1″ colspan=”1″ PMNS /th th valign=”bottom” align=”left” rowspan=”1″ colspan=”1″ EPI REAC /th th valign=”bottom” align=”left” rowspan=”1″ colspan=”1″ MEM /th th valign=”bottom” align=”left” rowspan=”1″ colspan=”1″ MES /th th valign=”bottom” align=”left” rowspan=”1″ colspan=”1″ SCLER /th /thead (+/+)30%30%60%60%40%10%(+/?)030%30%30%60%0(?/?)0020%000 Open in a separate window Kidneys from female NZM 2410 mice of various ER genotypes were scored 0C3+ for hypercellularity (HC), neutrophils/cell debris (PMNS), epithelial reactivity (EPI REAC), membranous changes (MEM), segmental mesangial expansion (MES), and sclerosis (SCLER). Ideals represent a share of 10 mice scored for every combined group. ER lacking and heterozygote females shown a nonsignificant tendency towards reduced glomerular pathology ratings in comparison to wildtype NZM 2410 mice (Fig. 2C). There have been no variations seen in renal ratings of NZM 2410 male mice, no matter ER position (Fig. 2D). IgG and C3 kidney deposition in ER lacking NZM2410 mice We following evaluated renal deposition of go with element C3 and IgG by immunofluorescence. Remarkably, regardless of the significant variations in albuminuria and pathology, there is no difference altogether IgG CPI-637 deposition between your three study organizations (Fig. 3A). When C3 deposition was evaluated (Fig. 3B), there is a minimal reduction in the ER lacking feminine mice in comparison to wildtype littermates that didn’t reach statistical significance. Man ER KO mice shown no difference in either IgG or C3 glomerular deposition in comparison to wildtype litter-mates (Figs. 3C and D). Numbers F and 3E display CPI-637 the diffuse glomerular staining seen in wildtype and ER KO females respectively. There have been no differences in C3 or IgG deposition in ER deficient females or males in comparison to wildtype littermates. Open in another window Shape 3 Renal immune system complicated deposition in ER lacking NZM2410 mice. Woman CPI-637 NZM2410.