Contactin associates with sodium channel Nav1.3 in native tissues and increases channel density at the cell surface.
The upregulation of voltage-gated sodium channel Na(v)1.3 has been linked to hyperexcitability of axotomized dorsal root ganglion (DRG) neurons, which underlies neuropathic pain. However, factors that regulate delivery of Na(v)1.3 to the cell surface are not known. Contactin/F3, a cell adhesion molecule, has been shown to interact with and enhance surface expression of sodium channels Na(v)1.2 and Na(v)1.9. In this study we show that contactin coimmunoprecipitates with Na(v)1.3 from postnatal day 0 rat brain where this channel is abundant, and from human embryonic kidney (HEK) 293 cells stably transfected with Na(v)1.3 (HEK-Na(v)1.3). Purified GST fusion proteins of the N and C termini of Na(v)1.3 pull down contactin from lysates of transfected HEK 293 cells. Transfection of HEK-Na(v)1.3 cells with contactin increases the amplitude of the current threefold without changing the biophysical properties of the channel. Enzymatic removal of contactin from the cell surface of cotransfected cells does not reduce the elevated levels of the Na(v)1.3 current. Finally, we show that, similar to Na(v)1.3, contactin is upregulated in axotomized DRG neurons and accumulates within the neuroma of transected sciatic nerve. We propose that the upregulation of contactin and its colocalization with Na(v)1.3 in axotomized DRG neurons may contribute to the hyper-excitablity of the injured neurons.
Shah
Bhaval S
BS
Department of Neurology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Rush
Anthony M
AM
Liu
Shujun
S
Tyrrell
Lynda
L
Black
Joel A
JA
Dib-Hajj
Sulayman D
SD
Waxman
Stephen G
SG
D000818:Animals; D001369:Axons / Q000737:chemistry; D001921:Brain / Q000196:embryology / Q000378:metabolism; D015816:Cell Adhesion Molecules, Neuronal / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D058966:Contactins; D004553:Electric Conductivity; D005727:Ganglia, Spinal / Q000166:cytology; D006801:Humans; D062552:NAV1.3 Voltage-Gated Sodium Channel; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D009475:Neurons, Afferent / Q000166:cytology / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015222:Sodium Channels / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*
D015816:Cell Adhesion Molecules, Neuronal; D058966:Contactins; D062552:NAV1.3 Voltage-Gated Sodium Channel; D009419:Nerve Tissue Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; C478638:Scn3a protein, rat; D015222:Sodium Channels