Sodium channel Nav1.6 accumulates at the site of infraorbital nerve injury.

BACKGROUND: Sodium channel (NaCh) expressions change following nerve and inflammatory lesions and this change may contribute to the activation of pain pathways. In a previous study we found a dramatic increase in the size and density of NaCh accumulations, and a remodeling of NaChs at intact and altered myelinated sites at a location just proximal to a combined partial axotomy and chromic suture lesion of the rat infraorbital nerve (ION) with the use of an antibody that identifies all NaCh isoforms. Here we evaluate the contribution of the major nodal NaCh isoform, Nav1.6, to this remodeling of NaChs following the same lesion. Sections of the ION from normal and ION lesioned subjects were double-stained with antibodies against Nav1.6 and caspr (contactin-associated protein; a paranodal protein to identify nodes of Ranvier) and then z-series of optically sectioned images were captured with a confocal microscope. ImageJ (NIH) software was used to quantify the average size and density of Nav1.6 accumulations, while additional single fiber analyses measured the axial length of the nodal gap, and the immunofluorescence intensity of Nav1.6 in nodes and of caspr in the paranodal region. RESULTS: The findings showed a significant increase in the average size and density of Nav1.6 accumulations in lesioned IONs when compared to normal IONs. The results of the single fiber analyses in caspr-identified typical nodes showed an increased axial length of the nodal gap, an increased immunofluorescence intensity of nodal Nav1.6 and a decreased immunofluorescence intensity of paranodal caspr in lesioned IONs when compared to normal IONs. In the lesioned IONs, Nav1.6 accumulations were also seen in association with altered caspr-relationships, such as heminodes. CONCLUSION: The results of the present study identify Nav1.6 as one isoform involved in the augmentation and remodeling of NaChs at nodal sites following a combined partial axotomy and chromic suture ION lesion. The augmentation of Nav1.6 may result from an alteration in axon-Schwann cell signaling mechanisms as suggested by changes in caspr expression. The changes identified in this study suggest that the participation of Nav1.6 should be considered when examining changes in the excitability of myelinated axons in neuropathic pain models.

Increased sodium channel immunofluorescence at myelinated and demyelinated sites following an inflammatory and partial axotomy lesion of the rat infraorbital nerve.

The localization of sodium channels (NaChs) change following nerve lesions and this change may contribute to the development of increased pain states. Here we examine the change in distribution of NaChs within the rat infraorbital nerve (ION) two weeks after a combined inflammatory/partial axotomy lesion that results in behavior showing increased sensitivity to mechanical stimuli. Sections from experimental and normal control IONs were double-stained for indirect immunofluorescence using an antibody that identifies all NaCh isoforms and caspr-antibody to identify nodes of Ranvier, and a confocal microscope z-series of optically sectioned images were then obtained. ImageJ (NIH) software was used to quantify the area of pixels showing maximum NaCh intensity within both caspr and non-caspr associated accumulations. Analysis showed that the lesioned IONs had many more split nodes, heminodes and caspr-negative "naked" accumulations, a significantly increased area of NaCh staining within typical nodes and "naked" accumulations, as well as an increased density and size of significant accumulations when compared to normal IONs. This study demonstrates a dramatic redistribution and increased immunofluorescence of NaChs especially at myelinated and demyelinated sites in fibers located just proximal to the lesion. The remodeling of NaChs seen in this study may represent an important event associated with the development of increased nerve excitability after lesions.