Single-cell mapping of the glycocalyx in the adult human and mouse ocular surface

The glycocalyx is a critical but often underappreciated modulator of cellular behavior. Its diversity across cell types within tissues remains poorly understood, but recent advances in single-cell profiling now enable more precise analysis of cell surface composition. Here, we applied single-cell glycan and RNA sequencing to profile glycocalyx diversity across human and mouse ocular surface cell types. Glycocalyx patterns effectively distinguished epithelial subtypes, with corneal epithelial cells enriched in complex and high-mannose N-glycans, conjunctival cells in fucosylated structures, and goblet cells in O-glycans. We also observed dynamic changes during epithelial maturation, marked by regulated shifts in sialic acid structures. In the mouse ocular surface, glycocalyx patterns distinguished major cell types, but the glycan profiles differed from those in humans, pointing to species-specific features. These findings demonstrate that glycocalyx composition is closely linked to cell identity and maturation and provide a foundation for exploring its roles in tissue organization and disease. Overall design: Single-cell RNA sequencing (scRNA-seq) analysis, utilizing nine distinct lectins conjugated to DNA, to characterize the glycocalyx composition of human and mouse ocular surface cells. The experiment consists of three scRNA-seq capture reactions per species, leveraging lectin-based profiling to reveal glycocalyx diversity