Alcohol withdrawal promotes anxiety-like behavior via activating glutaminergic neurons in the piriform cortex of male mice.

Alcohol withdrawal syndrome manifests clinically as anxiety, fatigue, emotional dysregulation, and sleep disturbances, significantly compromising both physical and psychological well-being. While these behavioral manifestations are known to involve limbic system dysfunction, the precise neural mechanisms underlying these effects remain poorly understood. In this study, we established a mouse model of alcohol withdrawal. Behavioral phenotypes were quantitatively assessed using the open field test (OFT) and elevated plus maze (EPM). The expression of c-fos in each brain region and NPY, PV, VGLuT1 neurons in (piriform cortex) PC were detected by immunofluorescence colocalization assay. Then VGLuT1 neurons in the PC was specifically inhibited or activated in VGLuT1-Cre mice and the behavioral changes were analyzed. The results indicated that mice exhibited anxiety-like behavior and weakened mobility after 3 days of alcohol withdrawal, accompanied by the increased activity of VGLuT1 neurons in the PC. Inhibiting or activating PC VGLuT1 neurons could decrease or enhance anxiety-like behavior and low mobility in mice, respectively. These findings indicate that 3 days of alcohol withdrawal activates the VGLuT1 neurons in the PC that can mediate decreased mobility and induce anxiety-like behaviors in mice.