OBJECTIVES: To investigate whether electroacupuncture (EA) at "Yanglingquan" (GV34) and "Xuanzhong" (GB34) can alleviate anxio-depression-like behaviors induced by neuropathic pain and whether the effect of EA is in connection with the activity of glutamatergic neurons in the ventrolateral orbital cortex (VLO). METHODS: The neuropathic pain model was established by common peroneal nerve ligation (CPNL). GV34 and GB34 were selected in EA treatment (2 Hz/15 Hz, 1 mA), 15 min each time, once a day for 7 d. Chemogenetic mixture viruses rAAV-VGLUT1-CRE-WPREs and rAAV-Ef1a-DIO-hM3Dq-mCherry-WPREs or rAAV-Ef1a-DIO-hM4Di-mCherry-WPREs were injected into the VLO, respectively. Clozapine-N-oxide (CNO) was injected intraperitoneally to regulate the activity of glutamatergic neurons in the VLO precisely. Mechanical withdrawal threshold (MWT) and paw withdrawal latency (PWL) were measured by dynamic plantar tactile instruments and plantar thermal nociceptor instruments. Anxio-depression-like behaviors were measured by behavioral tests, including open field test, elevated plus maze test, forced swimming test, and tail suspension test. The activation and inhibition levels of neurons were observed by immunofluorescence staining. RESULTS: Compared with the sham group, the MWT and PWL of CPNL mice were significantly decreased ( CONCLUSIONS: EA at GV34 and GB34 can alleviate anxio-depression-like behaviors induced by neuropathic pain through the modulation of glutamatergic neuron activity in the VLO.