Ss staining in NSC 697286 site proximal tubules. Although a lower IHC score was observed in the cortex of females on the AO diet (14) vs. REG diet (26.5) at 6 weeks (Table 2 and Fig. 4, panel 3 vs. 4), no difference was seen in males. By 20 weeks (16 weeks on the AO diet), eNOS expression in cortex showed a higher number of positively stained tubules and IHC score (Fig. 4, panel 1 vs. 2 in females). In contrast, males at 20 weeks showed lower numbers of positive tubules, and lower values for IHC and glomerular scores with AO supplementation (Fig. 4, panel 5 vs. 6). Sex differences of eNOS expression were also observed. Males had more eNOS positive stained tubules in cortex than females at 6 and 20 weeks (Fig. 4, panel 5 vs.1 and panel 7 vs. 3). At 20 weeks, this was associated with higher IHC and glomerular scores in cortex (Table 2). eNOS was also higher in males on the AO diet at 6 weeks, with a larger number of stained tubules and higher IHC score in both cortex and medulla (Table 2 and Fig. 4, panel 8 vs. 4 and panel 10 vs. 9). However, by 20 weeks on this diet the number of eNOS positive tubules was significantly higher for females in the cortex (Table 2 and Fig. 4, panel 2 vs. 6).Acta Histochem. Author manuscript; available in PMC 2017 March 01.Slyvka et al.PageIn cortex the number of eNOS positive tubules and IHC scores decreased with increasing age in females, but not males on REG diet (Table 2 and Fig. 4, panel 3 vs. 1). A decrease in these parameters was also seen in males on AO diet with age (Table 2 and Fig. 4, panel 8 vs. 6), while females maintained eNOS expression in GSK343 chemical information cortical tubules. The glomerular score increased with age in males on REG diet (Table 2 and Fig. 4 panel 5 vs. 7), but not in females. On the REG diet at 20 weeks eNOS staining was also detected in the tubulointerstitium. 3.3 nNOS In the present study, as shown by others (Maeda et al., 2003, Shin et al., 2000, Yabuki et al., 2006), nNOS was detected in tubules in both cortex and medulla of diabetic rodents, but not in glomeruli. We also detected nNOS in some collecting ducts. At 20 weeks males on the AO diet had higher numbers of nNOS positively stained tubules in cortex when compared with males on the REG diet (Table 3 and Fig. 5, panel 2 vs. 6). No diet differences were observed in females. At 6 weeks, males on both diets had more nNOS in kidneys than their female counterparts. They had greater numbers of nNOS positive stained tubules in cortex and medulla and higher IHC scores in cortex. (Table 3 and Fig. 5, panels 1 vs. 5, 3 vs. 7 and 8 vs. 9). Males on REG diet at 20 weeks showed an age related decrease in both nNOS positive tubular score and IHC score in both cortex and medulla compared to REG diet males at 6 weeks (Table 3 and Fig. 5, panels 1 vs. 2 and 3 vs. 4). 3.4 iNOS In agreement with the results of others (Cosenzi et al., 2002, Fujihara et al., 2002, Hohenstein et al., 2008, Kuloglu and Aydin, 2014, Ptilovanciv et al., 2013), iNOS was detected in renal tubules in both cortex and medulla (Figure 6). A greater number of iNOS positive tubules was detected in 6 week males on AO diet than on the REG diet in both cortex and medulla. Total IHC score was also higher in medulla (Table 4 and Fig. 6, panels 1 vs. 2 and 7 vs. 8). No diet differences were observed in females. At 6 weeks males on the AO diet had more iNOS-positive stained tubules in cortex than females (Table 4 and Fig. 6, panel 1 vs. 3). Scores for iNOS positively stained tubules decreased in males on AO die.Ss staining in proximal tubules. Although a lower IHC score was observed in the cortex of females on the AO diet (14) vs. REG diet (26.5) at 6 weeks (Table 2 and Fig. 4, panel 3 vs. 4), no difference was seen in males. By 20 weeks (16 weeks on the AO diet), eNOS expression in cortex showed a higher number of positively stained tubules and IHC score (Fig. 4, panel 1 vs. 2 in females). In contrast, males at 20 weeks showed lower numbers of positive tubules, and lower values for IHC and glomerular scores with AO supplementation (Fig. 4, panel 5 vs. 6). Sex differences of eNOS expression were also observed. Males had more eNOS positive stained tubules in cortex than females at 6 and 20 weeks (Fig. 4, panel 5 vs.1 and panel 7 vs. 3). At 20 weeks, this was associated with higher IHC and glomerular scores in cortex (Table 2). eNOS was also higher in males on the AO diet at 6 weeks, with a larger number of stained tubules and higher IHC score in both cortex and medulla (Table 2 and Fig. 4, panel 8 vs. 4 and panel 10 vs. 9). However, by 20 weeks on this diet the number of eNOS positive tubules was significantly higher for females in the cortex (Table 2 and Fig. 4, panel 2 vs. 6).Acta Histochem. Author manuscript; available in PMC 2017 March 01.Slyvka et al.PageIn cortex the number of eNOS positive tubules and IHC scores decreased with increasing age in females, but not males on REG diet (Table 2 and Fig. 4, panel 3 vs. 1). A decrease in these parameters was also seen in males on AO diet with age (Table 2 and Fig. 4, panel 8 vs. 6), while females maintained eNOS expression in cortical tubules. The glomerular score increased with age in males on REG diet (Table 2 and Fig. 4 panel 5 vs. 7), but not in females. On the REG diet at 20 weeks eNOS staining was also detected in the tubulointerstitium. 3.3 nNOS In the present study, as shown by others (Maeda et al., 2003, Shin et al., 2000, Yabuki et al., 2006), nNOS was detected in tubules in both cortex and medulla of diabetic rodents, but not in glomeruli. We also detected nNOS in some collecting ducts. At 20 weeks males on the AO diet had higher numbers of nNOS positively stained tubules in cortex when compared with males on the REG diet (Table 3 and Fig. 5, panel 2 vs. 6). No diet differences were observed in females. At 6 weeks, males on both diets had more nNOS in kidneys than their female counterparts. They had greater numbers of nNOS positive stained tubules in cortex and medulla and higher IHC scores in cortex. (Table 3 and Fig. 5, panels 1 vs. 5, 3 vs. 7 and 8 vs. 9). Males on REG diet at 20 weeks showed an age related decrease in both nNOS positive tubular score and IHC score in both cortex and medulla compared to REG diet males at 6 weeks (Table 3 and Fig. 5, panels 1 vs. 2 and 3 vs. 4). 3.4 iNOS In agreement with the results of others (Cosenzi et al., 2002, Fujihara et al., 2002, Hohenstein et al., 2008, Kuloglu and Aydin, 2014, Ptilovanciv et al., 2013), iNOS was detected in renal tubules in both cortex and medulla (Figure 6). A greater number of iNOS positive tubules was detected in 6 week males on AO diet than on the REG diet in both cortex and medulla. Total IHC score was also higher in medulla (Table 4 and Fig. 6, panels 1 vs. 2 and 7 vs. 8). No diet differences were observed in females. At 6 weeks males on the AO diet had more iNOS-positive stained tubules in cortex than females (Table 4 and Fig. 6, panel 1 vs. 3). Scores for iNOS positively stained tubules decreased in males on AO die.
