To study the mechanism of the L-type voltage-gated calcium channel (L-VGCC) involved in neuronal injury induced by brain ischemia and reperfusion (I/R), transient (15 min) brain ischemia was induced by four-vessel occlusion of Sprague-Dawley (SD) rats. Tyrosine phosphorylation of NR2A and interaction of NR2A with Src and Pyk2 in hippocampus induced by brain ischemia and reperfusion (I/R) were determined by immunoprecipitation and immunoblot(ting). Tyrosine phosphorylation of NR2A in hippocampus was enhanced after I/R. Interaction of NR2A with Src and Pyk2, tyrosine phosphorylation and kinase activity of Src and Pyk2 also increased after I/R. All the increases were partly inhibited by L-VGCC antagonist nifedipine administered to rats 20 min prior to brain ischemia. The results suggested that increase of tyrosine phosphorylation of NR2A induced by I/R had a relation to activation of L-VGCC. Src and Pyk2 interacting with NR2A might also be involved in this regulation of the tyrosine phosphorylation of NR2A induced by I/R.