Single-Cell RNA Sequencing of Rabbit Sclera at Different Developmental Stages: Unveiling Scleral Cells Atlas and the Heterogeneity of Fibroblasts

作者：Fayuan Li, Chengqi Gu, Chengpeng Liang, Li Yang, Shuo Wang, Qingqing Tang, Huan Jiang, Shaorong Linghu, Tingting Dan, Rong Shi, Xin Luo, Taixiang Liu · 发表于：Investigative Ophthalmology & Visual Science · 年份：2025 · DOI：10.1167/iovs.66.6.83 · 被引用次数：3 · 研究领域：Single-cell and spatial transcriptomics、Neurogenesis and neuroplasticity mechanisms、Barrier Structure and Function Studies

Purpose: This study aims to construct a single-cell transcriptomic atlas of the developing rabbit sclera to elucidate fibroblast heterogeneity, differentiation trajectories, matrisome expression patterns, and intercellular communication, while revealing conserved molecular features of scleral cells through cross-species analysis. Methods: Single-cell RNA sequencing (scRNA-seq) was performed on scleral tissues from New Zealand rabbits at embryonic day 25 (E25) and postnatal days 7 (P7), 21 (P21), and 180 (P180). Libraries were prepared using the DNBelab C Series Kit and sequenced on the BGISEQ-2000 platform. Sequencing reads were aligned to the OryCun2.0 genome using STAR, and unique molecular identifier (UMI) count matrices were generated with PISA. Data preprocessing was conducted using Seurat. Fibroblast lineage differentiation was analyzed via VIA, intercellular communication via CellChat, matrisome expression patterns via AUCell, and cross-species analyses via CACIMAR and hdWGCNA. Results: We identified 7 major cell types and 15 subpopulations, with fibroblasts dominating the cellular landscape. Distinct fibroblast subtypes exhibited varied expression profiles and functions: KERAlow SPARCL1⁺ fibroblasts showed stem/progenitor-like features, while KERAhigh myocilin (MYOC)⁺ fibroblasts displayed senescence-associated phenotypes. Matrisome analysis revealed dynamic alterations in collagen and extracellular matrix (ECM)-related genes, and intercellular communication analysis ...