title: Down-regulation of Stargazin inhibits the enhanced surface delivery of α-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor GluR1 subunit in rat dorsal horn and ameliorates postoperative pain.
abstract: BACKGROUND
  Stargazin is the first transmembrane protein known to regulate synaptic targeting of α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors. However, it is unclear whether regulation of the surface delivery of spinal AMPA receptor subunits by stargazin contributes to postoperative pain development.
  
  
  METHODS
  Western blot analysis was used to examine changes in the surface delivery of AMPA receptor subunits, GluR1 and GluR2, in rat dorsal horn. The interaction between stargazin and GluR1 and GluR2 was examined by coimmunoprecipitation. Expression of stargazin was suppressed by intrathecal administration of small interfering RNA311.
  
  
  RESULTS
  Membrane-bound GluR1, but not GluR2, in ipsilateral dorsal horn was increased at 3 h (1.49 ± 0.15-fold of β-tubulin, mean ± SEM) and 1 day (1.03 ± 0.25) after incision, as compared with that in control rats (naive, 0.63 ± 0.23, P < 0.05, n = 6 per group). The amount of GluR1 coimmunoprecipitated with stargazin was greater at 3 h after incision (1.48 ± 0.31-fold of input) than that in control animals (0.45 ± 0.24, P < 0.05, n = 6 per group). Importantly, the increase in membrane GluR1 at 3 h after incision was normalized to near control level (0.72 ± 0.20-fold of β-tubulin) by pretreatment with intrathecal stargazin small interfering RNA311 (0.87 ± 0.09), but not scrambled small interfering RNA (1.48 ± 0.24) or vehicle (1.25 ± 0.13, P < 0.05, n = 6 per group). Stargazin small interfering RNA311 pretreatment prevented the increase in stargazin-GluR1 interaction and decreased postoperative pain after incision.
  
  
  CONCLUSIONS
  This study suggests a critical role of stargazin-mediated surface delivery of GluR1 subunit in the development of postoperative pain. A better therapeutic strategy for postoperative pain may involve selectively down-regulating spinal stargazin to inhibit synaptic targeting of GluR1 subunit.
authors.affiliation: From the Department of Anesthesiology, Beijing Chao yang Hospital, Capital Medical University, Beijing, China (R.G., Y.Z., M.Z., Yun Wang, R.S., Y.L., J.X., A.W., Y.Y., Yun Wang); Department of Anesthesiology, Beijing Friendship Hospital, Capital Medical University, Beijing, China (R.G.); Department of Neurosurgery and Neurology, Lineberger Cancer Center, School of Medicine, University of North Carolina, Chapel Hill, North Carolina (J.W.); and Department of Anesthesiology and Critical Care Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland (Y.G.).
authors.forename: Ruijuan
authors.initials: R
authors.lastname: Guo
authors.forename: Yujie
authors.initials: Y
authors.lastname: Zhao
authors.forename: Meijuan
authors.initials: M
authors.lastname: Zhang
authors.forename: Yue
authors.initials: Y
authors.lastname: Wang
authors.forename: Rong
authors.initials: R
authors.lastname: Shi
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Jie
authors.initials: J
authors.lastname: Xu
authors.forename: Anshi
authors.initials: A
authors.lastname: Wu
authors.forename: Yun
authors.initials: Y
authors.lastname: Yue
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.forename: Yun
authors.initials: Y
authors.lastname: Guan
authors.forename: Yun
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D015536:Down-Regulation; D008297:Male; D010149:Postoperative Pain / Q000517:prevention & control*; D020671:Posterior Horn Cells / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D018091:Receptors, AMPA / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D002118:Calcium; D015220:Calcium Channels; D034741:RNA, Small Interfering; D018091:Receptors, AMPA; C491911:Cacng2 protein, rat; C104722:glutamate receptor ionotropic, AMPA 2
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