多组学揭示调控鸡子宫蛋壳强度的关键细胞类型与基因家族

Eggs represent an accessible and nutrient-dense source of high-quality animal protein, and decades of selective breeding have markedly elevated reproductive output in commercial laying hens. However, sustaining elevated productivity while improving eggshell integrity presents a critical challenge, as the molecular mechanisms of eggshell strength remain unclear. In this study, phenotypic assessment of eggshell strength was combined with single-cell transcriptomic profiling of the uterus from high- and low-strength groups, transcriptomic analysis of multiple tissues, and quantitative proteomic analysis of uterine fluid. Serum calcium and phosphorus levels did not differ significantly between groups. A single-cell atlas of the Rhode Island Red uterus was successfully generated for the first time, identifying nine distinct cell populations encompassing smooth muscle, epithelial, endothelial, and immune subsets. Integration of transcriptomic and proteomic datasets revealed that genes encoding collagens ( <i>COL4A1</i>/ <i>2</i>, <i>COL1A1</i>/ <i>2</i>, <i>COL5A1</i>, and <i>COL6A1</i>/ <i>2</i>/ <i>3</i>), solute carriers ( <i>SLC4A4</i>/ <i>7</i>, <i>SLC6A4</i>, <i>SLC9A2</i>/ <i>9</i>, and <i>SLC38A2</i>), ATPases ( <i>ATP1A1</i>, <i>ATP1B1</i>, <i>ATP2B1</i>/ <i>2</i>, <i>ATP2A2</i>/ <i>3</i>, and <i>ATP2C1</i>), calcium voltage-gated channels ( <i>CACNB2</i>, <i>CACNA1C</i>, and <i>CACNA2D1</i>), annexins ( <i>ANXA5</i> and <i>ANXA6</i>), and integrins ( <i>ITGB1</i> and <i>ITGA9</i>) were key molecular determinants associated with variation in eggshell strength. These genes were primarily enriched in signaling cascades involved in focal adhesion, actin cytoskeleton regulation, extracellular matrix (ECM)-receptor interactions, and calcium signaling. Notably, collagen family genes were predominantly localized to smooth muscle cells, consistent with the tissue remodeling and uterine inversion that occur during shell calcification, which may enhance spatial proximity between calcium ions and matrix proteins. These findings establish a multi-omics framework for understanding the uterine regulatory mechanisms underlying eggshell formation and offer a molecular foundation for breeding strategies aimed at prolonging laying cycles while preserving shell quality.