Figure 6.

ASIC1a contributes to pain-related increase in membrane trafficking of AMPAR GluA1 subunit in ACC. A, B, Representative images (A) and quantification (B) of Western blots showing that CFA injection did not induce significant changes in the expression of total ASIC1a in ACC (n = 5 mice per group). Ipsi, Ipsilateral; Contra, contralateral. C, D, Representative images (C) and quantification (D) of Western blots showing that CFA injection evoked a strong ASIC1a-independent upregulation of total GluN2B expression in ACC (WT-saline, n = 4 mice; WT-CFA, n = 4 mice; KO-saline, n = 4 mice; KO-CFA, n = 3 mice). E, F, Representative images (E) and quantification (F) of Western blots showing that CFA injection induced a significant increase in the membrane content of GluA1 and ASIC1a in ACC (WT-saline, n = 5 mice; WT-CFA, n = 5 mice; KO-saline, n = 4 mice; KO-CFA, n = 4 mice). The CFA-evoked membrane trafficking of GluA1 was not detected in ACC of ASIC1a KO mice. Membrane fractions were prepared by subcellular fractionation for assessing GluA1 and ASIC1a levels in the plasma membrane. G, H, Representative traces (G) and quantification (H) of evoked EPSCs demonstrating that CFA injection enhanced AMPAR-mediated synaptic transmission in ACC, which was attenuated by ASIC1a deletion (n = 8 neurons/4 mice). I–K, Representative traces (I) and quantification (J,K) of mEPSCs demonstrating that CFA injection resulted in increases in mEPSC frequency (J) and amplitude (K), and ASIC1a deletion blocked the change in mEPSC amplitude, but not frequency, in ACC neurons (n = 8 neurons/4 mice). L, M, Representative traces of paired-pulse ratio recording (L) and its quantification (M) demonstrating that ASIC1a is not involved in CFA-induced enhancement of presynaptic transmitter release (WT-saline, n = 10 neurons/5 mice; WT-CFA, n = 12 neurons/4 mice; KO-saline, n = 11 neurons/4 mice; KO-CFA, n = 12 neurons/5 mice). Data are mean ± SEM. *p < 0.05 versus saline. **p < 0.01 versus saline. ^#p < 0.05 versus WT, N.S., not significant.