Fyn kinase-mediated phosphorylation of NMDA receptor NR2B subunit at Tyr1472 is essential for maintenance of neuropathic pain.
Despite abundant evidence implicating the importance of N-methyl-D-aspartate (NMDA) receptors in the spinal cord for pain transmission, the signal transduction coupled to NMDA receptor activation is largely unknown for the neuropathic pain state that lasts over periods of weeks. To address this, we prepared mice with neuropathic pain by transection of spinal nerve L5. Wild-type, NR2A-deficient, and NR2D-deficient mice developed neuropathic pain; in addition, phosphorylation of NR2B subunits of NMDA receptors at Tyr1472 was observed in the superficial dorsal horn of the spinal cord 1 week after nerve injury. Neuropathic pain and NR2B phosphorylation at Tyr1472 were attenuated by the NR2B-selective antagonist CP-101,606 and disappeared in mice lacking Fyn kinase, a Src-family tyrosine kinase. Concomitant with the NR2B phosphorylation, an increase in neuronal nitric oxide synthase activity was visualized in the superficial dorsal horn of neuropathic pain mice by NADPH diaphorase histochemistry. Electron microscopy showed that the phosphorylated NR2B was localized at the postsynaptic density in the spinal cord of mice with neuropathic pain. Indomethacin, an inhibitor of prostaglandin (PG) synthesis, and PGE receptor subtype EP1-selective antagonist reduced the NR2B phosphorylation in these mice. Conversely, EP1-selective agonist stimulated Fyn kinase-dependent nitric oxide formation in the spinal cord. The present study demonstrates that Tyr1472 phosphorylation of NR2B subunits by Fyn kinase may have dual roles in the retention of NMDA receptors in the postsynaptic density and in activation of nitric oxide synthase, and suggests that PGE2 is involved in the maintenance of neuropathic pain via the EP1 subtype.
Abe
Tetsuya
T
Department of Medical Chemistry, Kansai Medical University, Moriguchi 570-8506, Japan.
Matsumura
Shinji
S
Katano
Tayo
T
Mabuchi
Tamaki
T
Takagi
Kunio
K
Xu
Li
L
Yamamoto
Akitsugu
A
Hattori
Kotaro
K
Yagi
Takeshi
T
Watanabe
Masahiko
M
Nakazawa
Takanobu
T
Yamamoto
Tadashi
T
Mishina
Masayoshi
M
Nakai
Yoshihide
Y
Ito
Seiji
S
D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects / Q000502:physiology; D015153:Blotting, Western; D015232:Dinoprostone / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D006651:Histocytochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D018345:Mice, Knockout; D016253:Microscopy, Immunoelectron; D009363:Neoplasm Proteins; D009569:Nitric Oxide / Q000378:metabolism; D010146:Pain / Q000209:etiology / Q000503:physiopathology* / Q000523:psychology; D010523:Peripheral Nervous System Diseases / Q000150:complications*; D010766:Phosphorylation; D010880:Piperidines / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology; D014443:Tyrosine / Q000378:metabolism; D019061:src-Family Kinases / Q000502:physiology*
C121001:NR2B NMDA receptor; D009363:Neoplasm Proteins; D010880:Piperidines; D016194:Receptors, N-Methyl-D-Aspartate; D009569:Nitric Oxide; D014443:Tyrosine; C095106:traxoprodil mesylate; D011505:Protein-Tyrosine Kinases; C490512:Frk protein, mouse; D019061:src-Family Kinases; D015232:Dinoprostone