Activation of Fibres in Rat Sciatic Nerve Alters Phosphorylation State of Connexin-43 at Astrocytic Gap Junctions in Spinal Cord: Evidence for Junction Regulation by Neuronal-glial Interactions

Intercellular communication via gap junction channels composed of connexin-43 is known to be regulated by phosphorylation of this protein. We investigated whether connexin-43 at astrocytic gap junctions is similarly regulated in response to neural activation. The effect of peripheral nerve stimulation on connexin-43 phosphorylation state in the spinal cord of rats was examined with a monoclonal antibody (designated 13-8300) shown previously to recognize selectively a dephosphorylated form of connexin-43. Immunolabelling with 13-8300 was absent in the lumbar spinal cord in control animals, but was induced in the dorsal horn ipsilateral to sciatic nerve electrical stimulation for 15min or 1h at a frequency of 1 or 100Hz. Immunorecognition of connexin-43 by a polyclonal anti-connexin-43 antibody, shown previously to undergo epitope masking under various conditions, was reduced in the dorsal horn on the stimulated side. These responses were abolished by local anaesthetic or tetrodotoxin application proximal to the site of nerve stimulation. Selective electrical stimulation of A-fibres or activation of cutaneous C-fibres by capsaicin evoked labelling with 13-8300 in deep and superficial laminae of the dorsal horn, respectively. Nerve stimulation increased the number of 13-8300-positive astrocytic gap junctions, as well as the levels of dephosphorylated connexin-43 in the dorsal horn on the stimulated side. Sciatic nerve transection produced results similar to those seen after C-fibre activation with capsaicin.Thus, peripheral nerve stimulation evokes astrocytic connexin-43 dephosphorylation in the spinal cord dorsal horn, suggesting that gap junctional coupling between astrocytes in vivo is subject to regulation by neuronal-glial interactions following neural activation.