A novel immune modulator IM33 mediates a glia-microbiota-neuronal axis that controls lifespan

Using Drosophila we identified a glia-secreted antimicrobial peptide IM33, which alters gut microbiota to modulate sleep. To understand how the gut commensal Lactiplantibacillus plantarum functions in the brain to regulate sleep, we performed single-cell RNA sequencing. Overall design: Single-cell RNA sequencing of fly brains from 3 age-matched groups: conventionally reared flies (CTRL), antibiotic-treated (8 days) flies (ABX) and Lactiplantibacillus plantarum-transferred (5-day ABX treatment + 3-day LP transfer) flies (LP). Briefly, 18 fly brains per group were dissected in cold Schneider's medium, followed by papain (100 unit/mL) and collagenase I (1.11 mg/mL) digestion. SYTOX-green negative cells are FACS sorted and utilized for sequencing.