Temporal and spatial patterns of Kv1.1 and Kv1.2 protein and gene expression in spinal cord white matter after acute and chronic spinal cord injury in rats: implications for axonal pathophysiology after neurotrauma.
After spinal cord injury (SCI), surviving white matter axons display axonal dysfunction associated with demyelination and altered K+ channel activity. To clarify the molecular basis of posttraumatic axonal pathophysiology after SCI, we investigated the changes in expression and distribution of the axonal K+ channel subunits Kv1.1 and Kv1.2 in spinal cord white matter after in vivo SCI in the rat. Using Western blot analysis, we found an increased expression of Kv1.1 and Kv1.2 at 2 and 6 weeks after SCI. By real-time PCR we observed an increase in Kv1.1 and Kv1.2 mRNA levels 1 day after SCI, which persisted until 6 weeks. Confocal immunohistochemistry showed a markedly dispersed labelling of Kv1.1 and Kv1.2 along the injured axons, in contrast to the tight localization of these channels to the juxtaparanodes of noninjured axons. This redistribution of Kv1.1 and Kv1.2 occurred as early as 1 h postinjury along some injured axons, and persisted at 6 weeks postinjury. In parallel with the redistribution of Kv1.1 and 1.2, contactin-associated protein (Caspr), which is normally confined to a paranodal location, also displayed a more diffuse distribution along the injured spinal cord axons. Our results suggest that the increased expression of Kv1.1 and Kv1.2 proteins is transcriptionally regulated. In contrast, the redistribution of the axonal K+ channel subunits occurs very early postinjury and probably reflects a disruption of the juxtaparanodal axonal region due to physical trauma, as shown by altered localization of Caspr.
Division of Neurosurgery, Toronto Western Research Institute, Krembil Neuroscience Center, University of Toronto, Toronto, Ontario, Canada M5T 2S8.
Soheila
S
Karimi-Abdolrezaee
Eftekhar
E
Eftekharpour
Michael G
MG
Fehlings
D000199:Actins / Q000378:metabolism; D000818:Animals; D001369:Axons / Q000378:metabolism; D015153:Blotting, Western; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism; D005260:Female; D015870:Gene Expression / Q000502:physiology; D007150:Immunohistochemistry / Q000379:methods; D051662:Kv1.1 Potassium Channel; D051663:Kv1.2 Potassium Channel; D007796:Laminectomy / Q000379:methods; D016900:Neurofilament Proteins / Q000378:metabolism; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism*; D024642:Potassium Channels, Voltage-Gated; D012333:RNA, Messenger; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013119:Spinal Cord Injuries / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D013194:Staining and Labeling; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism
D000199:Actins; D015816:Cell Adhesion Molecules, Neuronal; C105177:Cntnap1 protein, rat; C496972:Kcna2 protein, rat; D051663:Kv1.2 Potassium Channel; D016900:Neurofilament Proteins; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D012333:RNA, Messenger; D014404:Tubulin; C051774:neurofilament protein H; D051662:Kv1.1 Potassium Channel