Single-Cell Transcriptomic Profiling of the Mouse Conjunctiva Reveals Fibroblast-Driven Regulation, Immune Heterogeneity, and Neural Interactions

Despite its critical role in maintaining ocular surface integrity, the complex cellular heterogeneity and intercellular signaling mechanisms of the conjunctiva remain insufficiently characterized. In this study, we employed single-cell RNA sequencing (scRNA-seq) to systematically delineate the cellular composition and communication networks within the mouse conjunctiva. Conjunctival tissues obtained from 12- to 14-week-old wild-type C57BL/6J male mice were enzymatically dissociated and processed using the 10X Genomics Chromium platform. Rigorous quality control measures—including the exclusion of potential doublets and cells with elevated mitochondrial gene expression—were implemented prior to unsupervised clustering, which resolved eleven transcriptionally distinct cell populations, including fibroblasts, lymphoid cells, myeloid cells, Schwann cells, and endothelial cells. Differential gene expression analysis and intercellular communication inference via CellChat identified fibroblasts as central hubs orchestrating extensive cross-talk among diverse cell types. This high-resolution transcriptomic atlas of the mouse conjunctiva not only deepens our understanding of its intricate cellular landscape but also lays a critical foundation for future therapeutic strategies targeting ocular surface disorders.