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Time-Resolved Transcriptomic Analysis of Mammary Ductal Development, Lactation Function Activation, and Glandular Involution in Sows

ABSTRACT: To investigate the dynamic changes in gene expression during mammary gland development and lactation in sows, mammary tissue samples were collected from 30 multiparous crossbred half-sibling sows (Danish Landrace × Yorkshire) housed under standardized conditions. Samples were obtained at five critical physiological stages: mid-gestation (Day 70), late gestation (Day 110), early lactation (Day 2 postpartum), peak lactation (Day 10 postpartum), and early involution (Day 2 post-weaning), with six biological replicates per stage. Parenchymal mammary tissue (0.5–1 cm³) was excised from a defined location below the third pair of teats and rapidly snap-frozen in liquid nitrogen for RNA preservation. All instruments were treated with RNase inhibitors to ensure RNA integrity. The collected samples were stored at –80°C for subsequent total RNA extraction and transcriptome sequencing, providing a valuable resource for elucidating molecular mechanisms underlying mammary gland remodeling and lactation regulation in sows.

ORGANISM(S): Sus scrofa

PROVIDER: GSE295679 | GEO | 2026/05/20

REPOSITORIES: GEO

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Project description:The mammary gland is a dynamic tissue that undergoes most of its development postnatally to accommodate milk production during lactation and then, after weaning, undergoes postpartum involution, a coordinated process of epithelial cell death, remodeling, and inflammation that returns the tissue to a near pre-pregnant state. Yet, the mechanisms employed by some mammary cells to survive for subsequent lactation are not fully understood. Multiple studies suggest that involution is a driver of postpartum breast cancers (PPBCs), or those diagnosed within ten years of childbirth, that exhibit disproportionately high metastatic risk. Prior work implicates SEMA7A in mammary epithelial cell survival during involution and in metastasis of PPBC. We utilized single-cell RNA sequencing of mouse mammary glands during lactation and involution to delineate Sema7a-expressing populations during tissue development and remodeling. We found that Sema7a is expressed in a small subset of epithelial cells but is more predominantly expressed in endothelial and pericyte populations, whereas the developmentally regulated transcription factor Sox9 is highly expressed in epithelial subsets. We further show that SOX9 and SEMA7A expression changes across lactation and involution in mouse and in a unique set of human breast tissues collected from healthy donors during lactation and post-weaning. Using in vitro models of lactogenic differentiation and involution, we demonstrate a relationship between SOX9 and SEMA7A whereby knockdown of Sox9 in mammary epithelial cells increases SEMA7A expression and induces epithelial/mesenchymal hybrid phenotypes, identifying a potential regulatory axis in which SOX9 restrains SEMA7A-driven epithelial plasticity. Since SEMA7A is a driver of PPBC recurrence, we analyzed breast cancer datasets for SEMA7A and SOX9 to reveal that co-expression of SOX9 and SEMA7A correlates with metastatic risk in TNBC. Together, these findings define a previously unrecognized SOX9–SEMA7A relationship in the postpartum mammary gland and illustrate how studies of normal development can identify cellular mechanisms that may be susceptible to transformation and contribute to PPBC progression. This project aims to characterize transcriptomic changes in mouse mammary gland tissue across the reproductive stages of lactation and involution in wild-type mice. The goal is to understand cell-type-specific gene expression changes, with a focus on immune and epithelial compartments, and explore the potential role of SOX9 and SEMA7A in these processes.

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Time-Resolved Transcriptomic Analysis of Mammary Ductal Development, Lactation Function Activation, and Glandular Involution in Sows

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