High throughput transcriptomics analysis of ovine mammary epithelial cells stimulated with Staphylococcus aureus in vitro

Saif Adil Abbood Al-Janabi; Ghulam Asghar Sajid; Sidra Zeb; Muhammad Jasim Uddin; Mehmet Ulas Cinar

doi:10.1371/journal.pone.0333355

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High throughput transcriptomics analysis of ovine mammary epithelial cells stimulated with Staphylococcus aureus in vitro

Journal article

Open access

Peer reviewed

High throughput transcriptomics analysis of ovine mammary epithelial cells stimulated with Staphylococcus aureus in vitro

Saif Adil Abbood Al-Janabi, Ghulam Asghar Sajid, Sidra Zeb, Muhammad Jasim Uddin and Mehmet Ulas Cinar

PloS one, Vol.20(9), e0333355

2025

DOI: https://doi.org/10.1371/journal.pone.0333355

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Abstract

Animals

Annotations

Bioinformatics

Cattle

Cell activation

Cell culture

Cell injury

Cytoskeleton

Dairy industry

Epithelial cells

Epithelium

Gene expression

Genes

Genomes

Glucose

Health care

Immune response

Immune system

Innate immunity

Mammary gland

MAP kinase

Mastitis

Milk

NF-κB protein

Pathogenesis

Pathogens

Phagocytosis

Protein biosynthesis

Protein synthesis

Proteins

Real time

Ribonucleic acid

RNA

Sheep

Staphylococcus aureus

Staphylococcus infections

Stimulation

Transcriptomics

In sheep, the innate immune response of mammary epithelial cells (MECs) plays a central role in combating mastitis, yet our understanding of their resistance mechanisms remains limited. This study aimed to elucidate the gene expression profiles of ovine MECs following in vitro stimulation with Staphylococcus aureus (S. aureus) using RNA-Seq technology. Bioinformatics analysis identified a total of 175 differentially expressed genes (DEGs), including 172 up-regulated and 3 down-regulated genes in the stimulated group compared to the non-stimulated control group. Gene ontology annotation and functional pathway analysis indicated that these DEGs are primarily involved in ribosomal functions, which are essential for protein synthesis and first target of pathogens, as well as in immune response dysregulations, infection, phagocytosis, and bacterial invasion of epithelial cells. Validation via quantitative real-time PCR (qRT-PCR) confirmed the RNA-Seq results. Our results revealed that DEGs converged on innate immune pathways (TLR, NOD-like receptor, NF-κB, MAPK), cytoskeletal remodeling and translational control, indicating inflammatory activation and cell injury in oMECs and highlighting candidate targets for mastitis resistance selection against S. aureus. These findings significantly contribute to the understanding of how ovine MECs respond to S. aureus stimulation, providing a foundation for further research, particularly regarding the immune defense mechanisms, strategies and implications in dairy industry.

Details

Title: High throughput transcriptomics analysis of ovine mammary epithelial cells stimulated with Staphylococcus aureus in vitro
Authors/Creators: Saif Adil Abbood Al-Janabi - Erciyes University
Ghulam Asghar Sajid - Erciyes University
Sidra Zeb - Erciyes University
Muhammad Jasim Uddin - Murdoch University, Centre for Animal Production and Health
Mehmet Ulas Cinar
Publication Details: PloS one, Vol.20(9), e0333355
Publisher: Public Library of Science
Identifiers: 991005816849007891
Murdoch Affiliation: Harry Butler Institute; School of Veterinary Medicine; Centre for Animal Production and Health
Language: English
Resource Type: Journal article

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