NMDA receptor activation results in tyrosine phosphorylation of NMDA receptor subunit 2A(NR2A) and interaction of Pyk2 and Src with NR2A after transient cerebral ischemia and reperfusion.
Transient ischemia increases tyrosine phosphorylation of N-methyl-D-aspartate (NMDA) receptor. Several tyrosine kinases are involved in this process. In this study, effect of ischemia and reperfusion (I/R) on tyrosine phosphorylation of NMDA receptor subunit 2A (NR2A) and the interaction of two tyrosine kinases, Src and Pyk2, with NR2A was investigated. Four-vessel occlusion was used to produce transient (15 min) cerebral ischemia in SD rats. Tyrosine phosphorylation of NR2A in hippocampus was enhanced after 15 min of reperfusion and reached its peak level at 6 h of reperfusion. The increase sustained for at least 24 h. Src and Pyk2 co-immunoprecipitated with NR2A and the binding increased after I/R, which also reached a peak at 6 h of reperfusion. Besides, Src and Pyk2 were activated after I/R. These increases were prevented by ketamine, a selective NMDA receptor antagonist, which was administered to the SD rats 20 min before ischemia. Moreover, Src and Pyk2 coprecipitated with each other. These data show that NR2A, Src and Pyk2 might form a protein complex in vivo and the interaction suggests a possible mechanism of signal transduction in the postischemic hippocampus.
Liu
Y
Y
Research Center for Biochemistry and Molecular Biology, Xuzhou Medical College, 84 West Huai-hai Road, Xuzhou, 221002, Jiangsu, China.
Zhang
G
G
Gao
C
C
Hou
X
X
D000818:Animals; D002545:Brain Ischemia / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D051418:Focal Adhesion Kinase 2; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D007150:Immunohistochemistry; D007649:Ketamine / Q000494:pharmacology; D008297:Male; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D010766:Phosphorylation / Q000187:drug effects; D011505:Protein-Tyrosine Kinases / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015427:Reperfusion Injury / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D014443:Tyrosine / Q000187:drug effects* / Q000378:metabolism; D019061:src-Family Kinases / Q000187:drug effects* / Q000378:metabolism
D018691:Excitatory Amino Acid Antagonists; C120997:NR2A NMDA receptor; D016194:Receptors, N-Methyl-D-Aspartate; D014443:Tyrosine; D007649:Ketamine; D011505:Protein-Tyrosine Kinases; D051418:Focal Adhesion Kinase 2; C496054:Ptk2b protein, rat; D019061:src-Family Kinases