Expression of auxiliary beta subunits of sodium channels in primary afferent neurons and the effect of nerve injury.
Multiple voltage-gated sodium channels are the primary mediators of cell excitability. They are multimers that consist of the pore-forming alpha subunit and auxiliary beta subunits. Although ion permeability and voltage sensing are primarily determined by the alpha subunit, beta subunits are important modulators of sodium channel function. The purpose of this study was to assess the effect of axotomy on the expression of beta subunits (beta(1), beta(2) and beta(3)) and coexpression of Na(v)1.3 and beta(3) subunits in the dorsal root ganglion (DRG). We used sciatic nerve transection models or spared nerve injury (SNI) models in the rat. In reverse transcriptase-polymerase chain reaction analysis, there were no significant differences between contralateral and ipsilateral DRGs of beta(1) and beta(2) mRNA 3 days after axotomy. beta(3) mRNA expression in ipsilateral DRGs increased significantly compared with contralateral DRGs 3 days after axotomy. In in situ hybridization histochemistry, beta(1) mRNA was predominantly expressed in medium- to large-size neurons, whereas beta(2) mRNA was expressed in small- to large-size neurons. There were no significant differences in beta(1) and beta(2) mRNA between contralateral and ipsilateral DRGs 3 days after axotomy. In contrast, beta(3) mRNA was mainly expressed in small neurons and occasionally in medium- to large-size neurons, and beta(3) mRNA expression in small c-type neurons in ipsilateral DRGs was increased significantly compared with contralateral DRGs. We examined beta(3) mRNA expression with one of alpha subunits, Na(v)1.3-ir, in DRG neurons after axotomy using the double labeling method. We found a high percentage of coexpression in injured DRG neurons: 83.6+/-2.8% of neurons expressing beta(3) mRNA were labeled for Na(v)1.3-ir; 70.1+/-3.1% of Na(v)1.3-ir neurons expressed beta(3) mRNA. We also examined the expression of beta(3) mRNA in DRG neurons in the SNI model, a neuropathic pain model. We used activating transcription factor 3 to identify axotomized neurons, and found that beta(3) mRNA up-regulation occurred mainly in axotomized neurons in the neuropathic pain model. These data strongly suggest that beta(3) expression in injured DRG neurons following axotomy might be an important pathomechanism of post-nerve injury pain in primary sensory neurons.
Takahashi
N
N
Department of Anatomy and Neuroscience, Hyogo college of Medicine, 1-1 Mukogawa-cho, Nishinomiya City, Hyogo 663-8501, Japan.
Kikuchi
S
S
Dai
Y
Y
Kobayashi
K
K
Fukuoka
T
T
Noguchi
K
K
D051700:Activating Transcription Factor 3; D000818:Animals; D019771:Axotomy / Q000379:methods; D001522:Behavior, Animal; D015152:Blotting, Northern; D002452:Cell Count; D004195:Disease Models, Animal; D004210:Dissection / Q000379:methods; D007839:Functional Laterality; D005727:Ganglia, Spinal / Q000293:injuries / Q000378:metabolism; D006930:Hyperalgesia / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008026:Ligation / Q000379:methods; D008297:Male; D062552:NAV1.3 Voltage-Gated Sodium Channel; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009422:Nervous System Diseases / Q000378:metabolism*; D009475:Neurons, Afferent / Q000502:physiology*; D010543:Peroneal Nerve / Q000293:injuries / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012584:Sciatic Nerve / Q000293:injuries / Q000378:metabolism; D015222:Sodium Channels / Q000378:metabolism*; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism
D051700:Activating Transcription Factor 3; C497108:Atf3 protein, rat; D062552:NAV1.3 Voltage-Gated Sodium Channel; D009419:Nerve Tissue Proteins; D021122:Protein Subunits; D012333:RNA, Messenger; C478638:Scn3a protein, rat; D015222:Sodium Channels; D014157:Transcription Factors