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.

尽管结膜在维持眼表完整性中发挥关键作用，但其复杂的细胞异质性与细胞间信号传导机制仍未得到充分表征。本研究采用单细胞RNA测序（single-cell RNA sequencing，scRNA-seq）技术，系统解析了小鼠结膜的细胞组成与通讯网络。我们从12至14周龄的野生型C57BL/6J雄性小鼠中获取结膜组织，经酶解消化后使用10X Genomics Chromium平台完成样本制备与测序。在开展无监督聚类分析前，研究团队实施了严格的质控流程，包括排除潜在的双细胞及线粒体基因表达水平升高的细胞，最终成功解析出11个转录特征各异的细胞群，涵盖成纤维细胞、淋巴样细胞、髓系细胞、施万细胞及内皮细胞。通过差异基因表达分析及基于CellChat的细胞间通讯推断，研究发现成纤维细胞是介导多种细胞间广泛互作的核心枢纽。这份小鼠结膜的高分辨率转录组图谱，不仅深化了学界对其复杂细胞图谱的认知，更为未来靶向眼表疾病的治疗策略奠定了关键基础。