Dataset Information

Staphylococcus aureus-induced proteomic changes in the mammary tissue of rats: A TMT study

ABSTRACT: Staphylococcus aureus is one of the most important pathogens causing mastitis in dairy cows. The objective of this study was to establish a rat mastitis model induced by a S.aureus infection and to explore proteins that change in proteomes of mammary tissue in different udders states,which provided a better understanding of the host immune response to S. aureus mastitis. Six 72 h postpartum rats were randomly divided into two groups (n=3), with either 100 µL of PBS (blank group) or a S. aureus suspension containing 2×107 CFU·ml−1 (challenge group) infused into the mammary gland duct,respectively. After 24 h of infection, the rats were sacrificed, and mammary gland tissue was collected. A tandem mass tag (TMT)-based technology was applied to compare the proteomes of healthy and mastitic mammary tissue. Compared with the control group, the challenge group had 555 proteins with significant differences in expression, of which 428 were significantly up-regulated (FC>1.2 and p<0.05) and 127 were down-regulated (FC>0.83 and p<0.05 or p<0.01). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that up-regulated differentially significant expressed proteins (DSEPs) were associated with mainly immune responses,including integrin alpha M,inter-α-trypsin inhibitor heavy chain 4,alpha-2-macroglobulin.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Rattus Norvegicus (rat)

TISSUE(S): Stem Cell

DISEASE(S): Breast Cancer

SUBMITTER: Lirong Lirong

LAB HEAD: Beijing Key Laboratory of Dairy Cow Nutrition

PROVIDER: PXD017540 | Pride | 2020-05-27

REPOSITORIES: Pride

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			Action	DRS
	20190703_TMT_10_120min_Cf11HK_bjhb1_1.raw	Raw
	20190703_TMT_10_120min_Cf11HK_bjhb1_10.raw	Raw
	20190703_TMT_10_120min_Cf11HK_bjhb1_11.raw	Raw
	20190703_TMT_10_120min_Cf11HK_bjhb1_2.raw	Raw
	20190703_TMT_10_120min_Cf11HK_bjhb1_3.raw	Raw

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Staphylococcus aureus-induced proteomic changes in the mammary tissue of rats: A TMT-based study.

Cai Lirong L Tong Jinjin J Zhang Zhaonan Z Zhang Yonghong Y Jiang Linshu L Hou Xiaolin X Zhang Hua H

PloS one 20200504 5

Staphylococcus aureus is one of the most important pathogens causing mastitis in dairy cows. The objective of this study was to establish a rat model of mastitis induced by S. aureus infection and to explore changes in the proteomes of mammary tissue in different udder states, providing a better understanding of the host immune response to S. aureus mastitis. On day 3 post-partum, 6 rats were randomly divided into two groups (n = 3), with either 100 μL of PBS (blank group) or a S. aureus suspens ...[more]

PMID: 32365127

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Project description:Mastitis is a common disease that hinders the development of dairy industry and animal husbandry. It leads to the abuse of antibiotics, the emergence of super drug-resistant bacteria, and poses a great threat to human food health and safety. Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) are the most common pathogens of mastitis in dairy cows and usually cause subclinical or clinical mastitis. CircRNAs and N6-methyladenosine (m6A) play important roles in immunological diseases. However, the mechanisms by which m6A modifies circRNA in bovine mammary epithelial cells remain poorly understood. The aim of our study was to investigate m6A-modified circRNAs in bovine mammary epithelial cells (MAC-T cells) injured by S. aureus and E. coli. The profile of m6A-modified circRNA showed a total of 1599 m6A peaks within 1035 circRNAs in the control group, 35 peaks within 32 circRNAs in the S. aureus group, and 1016 peaks within 728 circRNAs in the E. coli group. Compared with the control group, 67 peaks within 63 circRNAs were significantly different in the S. aureus group, and 192 peaks within 137 circRNAs were significantly different in the E. coli group. Furthermore, we found the source genes of these differentially m6A-modified circRNAs in the S. aureus and E. coli groups with similar functions according to GO and KEGG analyses, which were mainly associated with cells injury, such as inflammation, apoptosis, and autophagy. CircRNA-miRNA-mRNA interaction networks predicted the potential circRNA regulation mechanism in S. aureus- and E. coli-induced cell injury. We found that the mRNAs in the networks, such as BCL2, MIF and TNFAIP8L2, greatly participated in the MAPK, WNT, and inflammation pathways. This is the first report on m6A-modified circRNA regulation of cells under S. aureus and E. coli treatment, and sheds new light on potential mechanisms and targets from the perspective of epigenetic modification in mastitis and other inflammatory diseases.

Project description:Establishment of an in vitro system to explore molecular mechanisms of mastitis susceptibility in cattle by comparative expression profiling of Escherichia coli and Staphylococcus aureus inoculated primary cells sampled from cows with different genetic predisposition for somatic cell score Primary bovine mammary gland epithelial cells (pbMEC) were sampled from the udder parenchyma of cows that were selected for high and low mastitis susceptibility by applying a marker assisted selection strategy considering QTL and molecular marker information of a repetitively confirmed QTL for SCS in the telomeric region on BTA18 The cells were cultivated and subsequently inoculated with heat inactivated mastitis pathogens Escherichia coli and Staphylococcus aureus, respectively. After 1, 6 and 24 hours the cells were harvested and comparatively analyzed using microarray expression chip technology to identify differences in mRNA expression profiles attributed to cultivation, inoculation or to genetic predisposition. Six heifers inheriting the favorable paternal QTL allele and five heifers inheriting the unfavorable QTL allele were selected by applying a marker assisted selection strategy. All heifers were kept under the same environmental conditions, had no clinical mastitis and did not show any indication of bacterial infection at slaughter. Primary bovine mammary gland epithelial cell cultures were established from cells sampled from the udder parenchyma of each cow. The cells were challenged with heat inactivated Escherichia coli and Staphylococcus aureus or with PBS for control. After 1, 6 and 24 hours the cells were harvested and mRNA expression was comparatively analyzed between time points for each treatment and each paternally inherited SCS-BTA18-QTL allele, respectively. In addition, the differences in gene expression at time points 1, 6 and 24h between inoculated and respective uninoculated control cells were investigated using the short time series expression miner STEM for co-expression profiling and GO cattegory enrichment analyses.