Figure 3

Alteration of expression of GABA _A receptor subunits and KCC2 in KO or KD mice. (A) Expression of GABA_A receptor subunits in PRIP-1 KO (open column) and DKO (close column) mice. Expression levels of PRIP-1 KO and DKO are based on the corresponding WT (represented as 100%, n = 5). *P < 0.05, for values in DKO vs PRIP-1 KO mice (Student’s t-test). (B) Expression of KCC2 in PRIP-1 KO (open column) and DKO (close column) mice. Expression levels represent each corresponding WT value as 100%. *P < 0.05, for values in DKO vs PRIP-1 KO mice (Student’s t-test). (C) Expression of GlyRα1 subunit, NKCC1 and KCC2 in WT and DKO 1 day after PSNL or sham operation (sham). The level of immunoreactivity was normalized to β-tubulin (mean ± S.E.M., n = 5–7). *P < 0.05, compared with values in WT sham-operated mice (Dunnet test). (D) R-DIOA administration induces pain relation behavior. Intrathecal administration of R-DIOA dose-dependently induced a decreased pain withdrawal threshold in WT mice. (E) Influence of KCC2 activity on pain sensitivity in PSNL mice. Each siRNA injection was performed 10 days after PSNL surgery. After 3 days of siRNA injection, R-DIOA (3 μg/mouse) was administrated intrathecally, and a paw withdrawal test was carried out 30 min after the injection. PSNL-operated WT mice were also treated with R-DIOA at 8 days after the surgery (representing as 8 in graph) and performed a paw withdrawal test. Naive represents prior to the surgery. Lines used in graph are as follows: black, green, violet, and blue are for WT, PRIP-1 KD, PRIP-2 KD, and DKD mice, respectively. The graph shows the withdrawal threshold value (mean ± S.E.M., n = 8). *P < 0.05, for values in 30 min after injection vs before injection (Student’s t-test).