In-situ glial cell-surface proteomics identifies pro-longevity factors in Drosophila

Much focus has shifted towards understanding how glial dysfunction contributes to age-related neurodegeneration due to the critical roles glial cells play in maintaining brain health. Cell-cell interactions, which are largely mediated by cell-surface proteins, control many critical aspects of development and physiology; as such, dysregulation of glial cell-surface proteins is hypothesized to play an important role in age-related neurodegeneration. However, it remains technically difficult to profile glial cell-surface proteins in intact brains. Here, we applied an in-situ cell-surface proteomic profiling method to glial cells from intact fly brains. Applying this platform to young and old flies, we identified candidate genes predicted to be involved in brain aging. Through a genetic screen, we identified one surface protein, DIP-β, which is down-regulated in old flies and can increase fly lifespan when overexpressed in adult glial cells. We performed whole-head single-nucleus RNA-seq and revealed that DIP-β overexpression mainly impacts glial and fat cells. We also found that glial DIP-β overexpression was associated with improved cell-cell communication. Our study is the first to apply in-situ cell-surface proteomics to glial cells in , and to identify DIP-β as a potential glial regulator of brain aging.