Different point stimulations can induce brain activity in specific regions, and however whether these stimulations affect unique neurotransmitter transmission remains unknown. Therefore, we aimed to investigate the effect of point specificity to the brain by resolving the metabolite profiles. Eighteen Sprague-Dawley rats were randomly divided into three groups: (1) the sham group: sham acupuncture at Zusanli (ST36) without electric stimulation; (2) the Zusanli (ST36) group: electroacupuncture (EA) at ST36; and (3) the Neiguan (PC6) group: EA at PC6. Then, the metabolites from rat brain samples were measured by LC-ESI-MS. The results of a partial least squares discriminant analysis revealed the differences among the sham, ST36, and PC6 groups regarding the relative content of metabolites in the cerebral cortex, hippocampus, and hypothalamus. EA at PC6 resulted in downregulation of adenosine, adrenaline, γ-aminobutyric acid, glycine, and glutamate majorly in hippocampus, and then in cerebral cortex. Otherwise, EA at ST6 resulted in upregulation of adrenaline and arginine in hippocampus, and all stimulations showed barely change of identified neurotransmitters in hypothalamus. These differential metabolite and neurotransmitter profiles prove that brain areas can be modulated by point specificity and may provide a maneuver to understand more details of meridian. Acupuncture stimulation can affect brain activity, including the somatosensory system, motor area, and limbic system; additionally, it can modulate the activity of specific brain areas. Hence, different points produce different effects, which is a phenomenon referred to as point specificity 1. Different points on the same meridian produce brain activity in similar area. For instance, acupuncture at the liver meridian Taichong (LR3) and Zhongdu (LR6) induces activity in the ipsilateral superior parietal lobe 2. Acupuncture at adjacent points LR3 and Neiting (ST44) at different meridians induces distinct cerebral activation patterns, and this accounts for the mechanism of point specific therapeutic effects 3. Overall, acupoints are specific to induce certain downstream modulation. A possible mechanism of acupuncture is the regulation of dopamine release to affect other neurotransmitters such as γ-aminobutyric acid (GABA), glutamate, serotonin, and opioids, which contribute to the balance of the central nervous system (CNS) 4. Acupuncture facilitated synaptic dopamine release in a mouse model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Parkinson's disease 5. Acupuncture, including manual acupuncture, electroacupuncture (EA), transcutaneous electrical nerve stimulation, and laser stimulation at PC6, can reduce the incidence of postoperative nausea and vomiting, which is a similar effect to that observed from the use of antiemetics 6. Acupuncture at PC6 modulates activity in the cardiovascular system by influencing the brain to secrete inhibitory neurotransmitters such as GABA, opioids, and 5-hydroxytryptamine to attenuate the activity of sympathetic n...