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Objectives: Menopause induces changes in neuronal transmission, leading to anxiety and depression. Changes in the brain's glutamate levels cause psychological behavior in postmenopausal women. Omega-3 has been studied to improve some of these behaviors., Methods: Twenty-four female Wistar rats were divided into four groups: sham-operated treated with water (SO-W), sham-operated treated with omega-3 (SO-O), ovariectomized (OVX) treated with water (OVX-W), and bilateral OVX treated with omega-3 (OVX-O). These treatments were performed for 20 days via gavage, before and after surgery, totaling 40 days., Results: In the forced swimming, elevated plus-maze, and open field tests to assess behaviors, such as depression and anxiety, omega-3 improved these behaviors in both treated groups. The levels of thiobarbituric acid reactive substances (TBARS) in the brain were not different between the groups; however, there was a significant decrease in the catalase activity in the SO-O group compared with the SO-W group ( P < 0.05). The glutamate level in the cerebrospinal fluid (CSF) was elevated in the SO-O group ( P < 0.001) but not in the OVX-W or OVX-O groups., Conclusions: These results bring novel data when related to the glutamatergic system in the SO-O group. This has suggested that the action mechanism of omega-3 was not dependent on glutamate levels in the CSF of the OVX group, but it played a regulatory role in the sham-operated animals. To confirm this, more studies are needed to explore this field when relating to the estrogen and glutamate receptor changes in specific brain regions., Competing Interests: No potential conflict of interest relevant to this article was reported., (Copyright © 2021 by The Korean Society of Menopause.)