In order to investigate whether excitatory neurotransmission system takes roles in tau phosphorylation caused by cold water stress, mice were treated with cold water stress (CWS), which were forced to swim at 4℃ for 5 min. The tau phosphorylation in mice brains was analyzed by immunobloting and immunohistochemistry with c-fos and phosphorylation-dependent tau antibodies. To evaluate the imbalance of excitatory or inhibitory neurotransmitters system, HPLC was used to detect amino acid neurotransmitters in brain after CWS. And the phosphorylated tau in brains of CWS mice, which were pre-treated with different antagonists for excitatory amino acid receptors and L-type calcium channel, was analyzed. The phosphorylated tau in hippocampus was significantly increased accompanied by an increase of c-fos expression at 1 h after CWS. HPLC showed that the content of all detected excitatory and inhibitory amino acid neurotransmitters appeared an acute increase then decrease pattern. At 15 min after CWS, aspartate and glycine appeared a significant increase, and aspartate, glutamate, taurine and GABA significantly decreased at 1 h. When the animals were pre-treated with NMDA receptor antagonist MK-801 (5 mg/kg) and AMPA receptor antagonist DNQX (0.5, 5 mg/kg), tau phosphorylation caused by CWS were significantly suppressed. Whereas, metabolic glutamate receptor antagonist, MAP-4, had no significant effect on tau phosphorylation. In addition, L-type calcium channel blocker, nimodipine (0.05, 0.5 mg/kg), also could inhibit CWS caused tau phosphorylation. These results indicated that CWS affects tau phosphorylation by mediating excitatory neurotransmission system through ionic excitatory amino acid receptors. The activation of excitatory neurotransmission system takes roles in CWS induced tau phosphorylation in hippocampus.