title: KATP channel subunits in rat dorsal root ganglia: alterations by painful axotomy.
abstract: BACKGROUND
  ATP-sensitive potassium (KATP) channels in neurons mediate neuroprotection, they regulate membrane excitability, and they control neurotransmitter release. Because loss of DRG neuronal KATP currents is involved in the pathophysiology of pain after peripheral nerve injury, we characterized the distribution of the KATP channel subunits in rat DRG, and determined their alterations by painful axotomy using RT-PCR, immunohistochemistry and electron microscopy.
  
  
  RESULTS
  PCR demonstrated Kir6.1, Kir6.2, SUR1 and SUR2 transcripts in control DRG neurons. Protein expression for all but Kir6.1 was confirmed by Western blots and immunohistochemistry. Immunostaining of these subunits was identified by fluorescent and confocal microscopy in plasmalemmal and nuclear membranes, in the cytosol, along the peripheral fibers, and in satellite glial cells. Kir6.2 co-localized with SUR1 subunits. Kir6.2, SUR1, and SUR2 subunits were identified in neuronal subpopulations, categorized by positive or negative NF200 or CGRP staining. KATP current recorded in excised patches was blocked by glybenclamide, but preincubation with antibody against SUR1 abolished this blocking effect of glybenclamide, confirming that the antibody targets the SUR1 protein in the neuronal plasmalemmal membrane. In the myelinated nerve fibers we observed anti-SUR1 immunostaining in regularly spaced funneled-shaped structures. These structures were identified by electron microscopy as Schmidt-Lanterman incisures (SLI) formed by the Schwann cells. Immunostaining against SUR1 and Kir6.2 colocalized with anti-Caspr at paranodal sites.DRG excised from rats made hyperalgesic by spinal nerve ligation exhibited similar staining against Kir6.2, SUR1 or SUR2 as DRG from controls, but showed decreased prevalence of SUR1 immunofluorescent NF200 positive neurons. In DRG and dorsal roots proximal to axotomy SLI were smaller and showed decreased SUR1 immunofluorescence.
  
  
  CONCLUSIONS
  We identified Kir6.2/SUR1 and Kir6.2/SUR2 KATP channels in rat DRG neuronal somata, peripheral nerve fibers, and glial satellite and Schwann cells, in both normal state and after painful nerve injury. This is the first report of KATP channels in paranodal sites adjacent to nodes of Ranvier and in the SLI of the Schwann cells. After painful axotomy KATP channels are downregulated in large, myelinated somata and also in SLI, which are also of smaller size compared to controls.Because KATP channels may have diverse functional roles in neurons and glia, further studies are needed to explore the potential of KATP channels as targets of therapies against neuropathic pain and neurodegeneration.
authors.affiliation: Department of Anesthesiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA. vzoga@mcw.edu
authors.forename: Vasiliki
authors.initials: V
authors.lastname: Zoga
authors.forename: Takashi
authors.initials: T
authors.lastname: Kawano
authors.forename: Mei-Ying
authors.initials: MY
authors.lastname: Liang
authors.forename: Martin
authors.initials: M
authors.lastname: Bienengraeber
authors.forename: Dorothee
authors.initials: D
authors.lastname: Weihrauch
authors.forename: Bruce
authors.initials: B
authors.lastname: McCallum
authors.forename: Geza
authors.initials: G
authors.lastname: Gemes
authors.forename: Quinn
authors.initials: Q
authors.lastname: Hogan
authors.forename: Constantine
authors.initials: C
authors.lastname: Sarantopoulos
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D000818:Animals; D019771:Axotomy / Q000009:adverse effects; D015740:Calcitonin Gene-Related Peptide / Q000378:metabolism; D005727:Ganglia, Spinal / Q000378:metabolism* / Q000648:ultrastructure; D005905:Glyburide / Q000494:pharmacology; D007004:Hypoglycemic Agents / Q000494:pharmacology; D007150:Immunohistochemistry; D054086:KATP Channels / Q000235:genetics / Q000378:metabolism*; D008297:Male; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D009437:Neuralgia / Q000209:etiology / Q000378:metabolism* / Q000503:physiopathology; D016900:Neurofilament Proteins / Q000378:metabolism; D010523:Peripheral Nervous System Diseases / Q000209:etiology / Q000378:metabolism* / Q000503:physiopathology; D024661:Potassium Channels, Inwardly Rectifying / Q000235:genetics / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D011901:Ranvier's Nodes / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D011955:Receptors, Drug / Q000235:genetics / Q000378:metabolism; D012583:Schwann Cells / Q000378:metabolism / Q000648:ultrastructure; D011984:Sensory Receptor Cells / Q000378:metabolism* / Q000648:ultrastructure; D064233:Sulfonylurea Receptors; D000099161:Kcnj11 Channel
chemical_list: D018528:ATP-Binding Cassette Transporters; D015740:Calcitonin Gene-Related Peptide; D005905:Glyburide; D007004:Hypoglycemic Agents; D054086:KATP Channels; D016900:Neurofilament Proteins; D024661:Potassium Channels, Inwardly Rectifying; D021122:Protein Subunits; D011955:Receptors, Drug; D064233:Sulfonylurea Receptors; D000099161:Kcnj11 Channel; C577509:Abcc8 protein, rat; C577512:Abcc9 protein, rat; C092520:uK-ATP-1 potassium channel; C051774:neurofilament protein H