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OBJECTIVE: To investigate the optimal timing and underlying mechanism of electroacupuncture(EA)at Baihui(GV 20)and Dazhui(GV 14)for improved long-term functional recovery after focal cerebral ischemia in a photothrombotic stroke mouse model.METHODS: Totally 50 adult male C57BL/6J mice were assigned into 5 groups:(a)the control group,sham-operated mice(n = 10);(b)the vehicle group,focal cerebral ischemia induction without EA(n = 10);(c)the acute EA group,mice received EA imme-diately post-ischemia,followed by once-daily treat-ments for 7 consecutive days(n = 10);(d)the sub-acute EA group,mice received EA 4 days post-isch-emia,followed by once-daily treatments for 7 con-secutive days(n = 10);(e)the delayed EA group.EA stimulation(2 Hz,2 V for 20 min)was applied to acupuncture points(acupoints),Baihui(GV 20)and Dazhui(GV 14),once a day for 7 consecutive days beginning immediately(acute treatment),4 d(sub-acute treatment)and 10 d(delayed treatment)af-ter focal cerebral ischemia in C57BL/6J mice.Behav-ioral OBJECTIVE: To investigate the optimal timing and underlying mechanism of electroacupuncture(EA)at Baihui(GV 20)and Dazhui(GV 14)for improved long-term functional recovery after focal cerebral ischemia in a photothrombotic stroke mouse model.METHODS: Totally 50 adult male C57BL/6J mice were assigned into 5 groups:(a)the control group,sham-operated mice(n = 10);(b)the vehicle group,focal cerebral ischemia induction without EA(n = 10);(c)the acute EA group,mice received EA immediately post-ischemia,followed by once-daily treatments for 7 consecutive days(n = 10);(d)the sub-acute EA group,mice received EA 4 days post-isch-emia,followed by once-daily treatments for 7 consecutive days(n = 10);(e)the delayed EA group.EA stimulation(2 Hz,2 V for 20 min)was applied to acupuncture points(acupoints),Baihui(GV 20)and Dazhui(GV 14),once a day for 7 consecutive days beginning immediately(acute treatment),4 d(sub-acute treatment)and 10 d(delayed treatment)after focal cerebral ischemia in C57BL/6J mice.Behavioral assessments were conducted 21 and 28 d post-ischemia and histopathological analyses were performed 28 days post-ischemia.RESULTS: The subacute EA treatment at Baihui(GV 20)and Dazhui(GV 14)significantly improved functional recovery compared to the vehicle group 28 d after ischemic brain injury,although brain atrophy was not reduced.The number of NeuN* and NeuN+/BrdU+cells as well as GFAP intensity in the ipsilateral cortex were significantly increased in the sub-acute group compared to the vehicle group 28 d post-ischemia.We concluded that EA stimulation 4 d post-ischemia(subacute treatment)enhanced neu-rogenesis and astrogliosis,likely contributing to long-term functional recovery following focal cere-bral ischemia.CONCLUSION: Our findings suggest that the tim-ing of the EA therapy at Baihui(GV 20)and Dazhui(GV 14)determines the therapeutic effects in mice with focal cerebral ischemia induced by photo-thrombotic occlusion.