Article Figures & Data
Figures
- Figure 1.
ECl- remains depolarized in Fmr1 ko mice during cortical development. A, Representative example of a perforated patch-clamp recording from a layer IV neuron in the somatosensory cortex of a P10 Fmr1 wt mouse. Recordings were made at several hold potentials and the ECl- calculated from the linear fit of the current–voltage relationship. GABA responses shown at −80, 0, and +40 mV were evoked by extracellular stimulation in the presence of glutamate blockers d-APV (50 μm) and CNQX (10 μm). Calibration for current traces: 50 ms, 200 pA. B, Representative recording from Fmr1 ko at P10 and current–voltage relationship of GABA-mediated currents. ECl- is significantly more depolarized at this age in recordings from Fmr1 ko mice. Calibration for current traces: 50 ms, 50 pA. C, Grouped data from all recordings. The average ECl- calculated from each individual recording is plotted against the age of the mouse (postnatal day). The RMP measured at P10 is denoted by the dashed line and shaded area represents points at which GABA would have a mature hyperpolarizing response. *p < 0.05 (P5: wt, n = 13; ko n = 4. P6: wt, n = 6; ko, n = 11. P7: wt, n = 6; ko, n = 4. P8: wt, n = 10: ko, n = 8. P9: wt, n = 8; ko, n = 11.P10: wt, n = 8; ko, n = 5. P11: wt, n = 16; ko, n = 14. P12: wt, n = 17; ko, n = 6. P13: wt, n = 12: ko, n = 5. P14: wt, n = 11: ko, n = 10. P15: wt, n = 12; ko, n = 7).
- Figure 2.
Expression of the juvenile chloride cotransporter NKCC1. A, The expression of NKCC1 is elevated in the cortex of Fmr1 ko mice at the close of the critical period. Representative gel run with cortical homogenates from Fmr1 wt and Fmr1 ko mice and probed with anti-NKCC1 and anti-β-tubulin (β-tub). Western blots were used to measure NKCC1 protein levels at three developmental time points: P5, P10, and P15. B, Grouped data from all experiments. The average intensity of NKCC1 was normalized to β-tubulin in Fmr1 wt and Fmr1 ko in each sample at each age (P5: wt, n = 8; ko, n = 8. P10: wt, n = 11; ko, n = 11. P15: wt, n = 8; ko, n = 7). A significant elevation of NKCC1 protein levels was observed at P10 in samples from the Fmr1 ko mice. *p < 0.05. C, Representative Western blots for cortical homogenates probed with anti-KCC2 antibodies from Fmr1 wt and Fmr1 ko mice at P5, P10, and P15. D, Grouped data for all KCC2 Western blots (P5: wt, n = 8; ko, n = 8. P10: wt, n = 11; ko, n = 11; P15: wt, n = 8; ko, n = 7). No difference in relative protein levels was observed at any age between the two genotypes (p > 0.05). All values are means ± SEM.
- Figure 3.
Chloride cotransporter transcript expression is not altered in cortex of Fmr1 ko mice. A, Relative transcript abundance of NKCC1 in cortex of Fmr1 wt and Fmr1 ko at P5, P10, and P15. B, Relative RNA expression of KCC2 in Fmr1 wt and Fmr1 ko at P5, P10, and P15. Rq was calculated as the ratio of the target gene to GAPDH as the reference gene (p > 0.05 for all age groups for both NKCC1 and KCC2).
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