Activation of G protein gated inwardly rectifying potassium (GIRK) channels in keratinocytes mediates peripheral kappa opioid receptor-mediated antinociception

Kappa opioid receptors (KOR) expressed by peripheral pain-sensing neurons (nociceptors) are a promising target for development of effective and safer analgesics for inflammatory pain that are devoid of central nervous system adverse effects. Here we sought to delineate the signaling pathways that underlie peripheral KOR-mediated antinociception in adult male and female Sprague-Dawley rats. In an inflammatory model of pain, local intraplantar (i.pl.) injection of pertussis toxin prevented antinociception induced by the KOR agonist, U50488, indicating that members of the Gi/o family mediate the antinociceptive response. Furthermore, i.pl. injection of the G protein-coupled inward-rectifying potassium (GIRK) channel blocker, TPNQ, as well as GIRK2 subunittargeted siRNA abolished U50488-mediated antinociceptive behavioral responses in both male and female rats. Consistent with these data, i.pl. injection of ML297, a direct activator of GIRK1 subunit-containing channels, elicited peripheral antinociceptive behavior. It is well known that intraepidermal nerve fibers (IENF) that innervate the hindpaw propagate nociceptive signals to the spinal cord. However, recent studies suggest that keratinocytes, the major cell type in the epidermis, also play an active role in pain and sensory processing. Results from RT-qPCR, RNAscope and immunohistochemistry experiments confirmed that both KOR and GIRK are expressed in keratinocytes in the epidermal layer of the rat hindpaw. Knockdown of either KOR or GIRK2 subunits selectively in keratinocytes by i.pl. injection of shRNA plasmids, prevented the antinociceptive response to U50488. Taken together, these data suggest that KOR-mediated activation of GIRK channels in keratinocytes is required for peripherally-mediated antinociception.