Down-regulation of K+–Cl− co-transporter 2 in mouse medullary dorsal horn contributes to the formalin-induced inflammatory orofacial pain

Cation chloride co-transporters, including K+–Cl− co-transporter 2 (KCC2) and Na+–K+–Cl− co-transporter 1 (NKCC1), are of particular importance to GABAergic transmission and thus involved in the development of hyperalgesia at the spinal level. However, there are fewer relevant reports in the trigeminal system. In this study, we investigated the behavioral changes and the accompanying change in the expressions of KCC2 and NKCC1 mRNAs in mouse medullary dorsal horn (MDH) after subcutaneous injection of formalin into the left vibrissa pad. Furthermore, we observed the behavioral changes following intracisternal injection of KCC2 antisense oligodeoxynucleotides (ASO) into naïve mice. Subcutaneous injection of formalin-induced a significant increase in the number of face rubbing events which are the indicators of spontaneous pain. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) results indicated that, compared to the saline control or the contralateral side, the ipsilateral mRNA level of KCC2 but not NKCC1, was significantly reduced in formalin-injected mice during phase 1 observation, followed by gradual recovery. Intracisternal injection of KCC2 ASO into naïve mice led to behavioral hypersensitivity similar to the hyperalgesia observed in formalin experiments. These findings indicate that peripheral inflammation induces down-regulation of KCC2 in the MDH, which may in turn facilitate the development of acute inflammatory pain. These results also suggest that preventing the down-regulation of KCC2 is a possible way to combat orofacial pain.