title: Spinal serum-inducible and glucocorticoid-inducible kinase 1 mediates neuropathic pain via kalirin and downstream PSD-95-dependent NR2B phosphorylation in rats.
abstract: The coupling of the spinal postsynaptic density-95 (PSD-95) with the glutamatergic N-methyl-d-aspartate receptor NR2B subunit and the subsequent NR2B phosphorylation contribute to pain-related plasticity. Increasing evidence reveals that kalirin, a Rho-guanine nucleotide exchange factor, modulates PSD-95-NR2B-dependent neuroplasticity. Our laboratory recently demonstrated that serum-inducible and glucocorticoid-inducible kinase 1 (SGK1) participates in inflammation-associated pain hypersensitivity by modulating spinal glutamatergic neurotransmission. Because kalirin is one of the proteins in PSD that is highly phosphorylated by various kinases, we tested whether kalirin could be a downstream target of spinal SGK1 that participates in neuropathic pain development via regulation of the PSD-95-NR2B coupling-dependent phosphorylation of NR2B. We observed that spinal nerve ligation (SNL, L5) in male Sprague Dawley rats resulted in behavioral allodynia, which was associated with phosphorylated SGK1 (pSGK1), kalirin, and phosphorylated NR2B (pNR2B) expression and an increase in pSGK1-kalirin-PSD-95-pNR2B coprecipitation in the ipsilateral dorsal horn (L4-L5). SNL-enhanced kalirin immunofluorescence was coincident with pSGK1, PSD-95, and pNR2B immunoreactivity. Small-interfering RNA (siRNA) that targeted spinal kalirin mRNA expression (10 μg, 10 μl; i.t.) reduced SNL-induced allodynia, kalirin and pNR2B expression, as well as kalirin-PSD-95 and PSD-95-pNR2B coupling and costaining without affecting SGK1 phosphorylation. Daily administration of GSK-650394 (an SGK1 antagonist; 100 nm, 10 μl, i.t.) not only exhibited effects similar to the kalirin mRNA-targeting siRNA but also attenuated pSGK1-kalirin costaining and SGK1-kalirin coupling. We suggest that nerve injury could induce spinal SGK1 phosphorylation that subsequently interacts with and upregulates kalirin to participate in neuropathic pain development via PSD-95-NR2B coupling-dependent NR2B phosphorylation.
authors.affiliation: Department of Medicine, Mackay Medical College, New Taipei 25160, Taiwan. hsien.yu@gmail.com
authors.forename: Hsien-Yu
authors.initials: HY
authors.lastname: Peng
authors.forename: Gin-Den
authors.initials: GD
authors.lastname: Chen
authors.forename: Cheng-Yuan
authors.initials: CY
authors.lastname: Lai
authors.forename: Ming-Chun
authors.initials: MC
authors.lastname: Hsieh
authors.forename: Tzer-Bin
authors.initials: TB
authors.lastname: Lin
mesh_terms: D000818:Animals; D001565:Benzoates / Q000494:pharmacology; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D000076122:Disks Large Homolog 4 Protein; D020662:Guanine Nucleotide Exchange Factors / Q000235:genetics / Q000378:metabolism*; D006930:Hyperalgesia / Q000378:metabolism / Q000503:physiopathology; D017874:Immediate-Early Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D008026:Ligation; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D009437:Neuralgia / Q000378:metabolism* / Q000503:physiopathology; D009473:Neuronal Plasticity / Q000502:physiology; D059348:Peripheral Nerve Injuries / Q000378:metabolism / Q000503:physiopathology; D010766:Phosphorylation / Q000502:physiology; D020671:Posterior Horn Cells / Q000378:metabolism / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism*; D013127:Spinal Nerves / Q000378:metabolism / Q000503:physiopathology; D013569:Synapses / Q000378:metabolism; D000099184:Serum-Glucocorticoid Regulated Kinases
chemical_list: D001565:Benzoates; D019086:Bridged Bicyclo Compounds, Heterocyclic; D000076122:Disks Large Homolog 4 Protein; D020662:Guanine Nucleotide Exchange Factors; D017874:Immediate-Early Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D017346:Protein Serine-Threonine Kinases; D034741:RNA, Small Interfering; D016194:Receptors, N-Methyl-D-Aspartate; D000099184:Serum-Glucocorticoid Regulated Kinases; C501449:Dlg4 protein, rat; C102365:Kalrn protein, rat; C121001:NR2B NMDA receptor; C532254:2-cyclopentyl-4-(5-phenyl-1H-pyrrolo(2,3-b)pyridin-3-yl)-benzoic acid