Figure 4
Perinatal testosterone limits KCC2 localisation at the membrane. (A) Western blot membrane of total KCC2 (images were cropped from the same gel). (B,C) Quantification of total KCC2 expression levels (monomer band at 140 kDa) in the hippocampus of different experimental groups do not show significant differences at P7 (B) (F: 1.28 ± 0.17 au, n = 8 rats from 3 litters; M: 1.18 ± 0.19 au, n = 8 rats from 3 litters; A: 1.19 ± 0.15 au, n = 8 rats from 3 litters; TFM: 1.38 ± 0.22 au, n = 8 rats from 3 litters; One-way ANOVA with Tukey’s post hoc analysis, p = 0.926) or at P40 (C) (F: 0.90 ± 0.12 au, n = 8 rats from 3 litters; M: 0.67 ± 0.11 au, n = 8 rats from 3 litters; A: 0.66 ± 0.09 au, n = 8 rats from 3 litters; TFM: 0.67 ± 0.09 au, n = 8 rats from 3 litters; One-way ANOVA with Tukey’s post hoc analysis, , p = 0.355). (D) Western blot membrane of KCC2 in membrane fractions (images were cropped from different gels). (E,F) Quantification of KCC2 monomer expression levels in the membrane fractions show sex differences in the hippocampus at P7 (E) (F: 1.07 ± 0.10 au, n = 12 rats from 4 litters; M: 0.48 ± 0.03 au, n = 12 rats from 5 litters; A: 0.58 ± 0.05 au, n = 8 rats from 3 litters; TFM: 1.10 ± 0.04 au, n = 5 rats from 3 litters; one-way ANOVA with Tukey’s post hoc analysis, F × M: p < 0.001; F × A: p = 0.001, TFM × M: p < 0.001; TFM × A: p = 0.001). Conversely, no sex differences were found at P40 (F) (F: 0.62 ± 0.08 au, n = 8 rats from 3 litters; M: 0.60 ± 0.04 au, n = 8 rats from 4 litters, A: 0.69 ± 0.11 au, n = 8 rats from 3 litters; TFM: 0.87 ± 0.06 au, n = 8 rats from 3 litters; One-way ANOVA with Tukey’s post hoc analysis, p = 0.121). Each lane represents a different animal. Blot shows representative samples of different sex groups. Dots represent individual data points. F females, M males, A andro/testosterone-treated females, T TFM/testosterone-insensitive males. Graphs represent mean ± SEM.
