Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Gene-expression profile of the response to E. coli (k2bh2) infection in the bovine udder

ABSTRACT: Mastitis in dairy cows is one of the most costly and prevalent diseases affecting dairy cows world wide. Insight in the molecular regulation of the host immune response to an E. coli infection, could help to develop new strategies to prevent cattle from E. coli infection. Here we performed a gene-expression analysis from udder tissue exposed to a controlled E. coli infection at T=24h post infection (p.i.) representing the acute phase response and at T=192h p.i. representing a chronic stage. 49 Samples

ORGANISM(S): Bos taurus

SUBMITTER: A.J. (Bart) Buitenhuis

PROVIDER: E-GEOD-24217 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

In depth analysis of genes and pathways of the mammary gland involved in the pathogenesis of bovine Escherichia coli-mastitis.

Buitenhuis Bart B Røntved Christine M CM Edwards Stefan M SM Ingvartsen Klaus L KL Sørensen Peter P

BMC genomics 20110228

<h4>Background</h4>Bovine mastitis is one of the most costly and prevalent diseases affecting dairy cows worldwide. In order to develop new strategies to prevent Escherichia coli-induced mastitis, a detailed understanding of the molecular mechanisms underlying the host immune response to an E. coli infection is necessary. To this end, we performed a global gene-expression analysis of mammary gland tissue collected from dairy cows that had been exposed to a controlled E. coli infection. Biopsy sa ...[more]

PMID: 21352611

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Project description:Once daily milking reduces milk yield and alters mammary transcriptome resulting in a decrease in milk protein synthesis as well as an induction of the apoptotic signaling networks. A local regulation due to milk stasis in the tissue could contribute to this effect but such mechanisms have not yet been described. To challenge this hypothesis, cows were milked unilaterally, once daily on one udder half and twice daily on the other one, and variations in gene expression were studied in biopsies as well as in mammary epithelial cells (MEC) shed into milk during the lactation process (milk MEC). This study therefore also contributes to decipher if transcript variations in milk purified MEC can reflect that of the mammary tissue. We compared the mammary transcript profiles in biopsies collected from unilaterally once versus twice-daily milked udder halves, 504 transcripts were differentially expressed: 193 and 232 transcripts were up- and down-regulated, respectively. A first category of transcripts, which accumulation levels are mostly up-regulated, relates to mechanisms involved in cell renewal, such as cell cycle, cellular growth and proliferation, cell death and cellular development. A second family, mostly down-regulated, is involved in small molecule biochemistry, amino acid, lipid and carbohydrate metabolisms as well as molecular transport. A third category, mostly up-regulated, includes transcripts expressed in non-epithelial mammary cells such as adipocytes, endothelial and immune cells and cells from the connective tissue. These results are consistent with previous data showing a decrease in mammary synthesis activity and an activation of cell death during once daily milking. They further suggest that during once daily milking the local milk accumulation has a major effect on mammary remodeling. Interestingly, some transcripts belonged to a third family described as M-BM-+ molecular transports M-BM-;. The expression of the 21 mRNA was then analyzed by RT-par in MEC (Table 4). Seven transcripts (NUCB2, RNASE4, ABCG2, RNASE1, SLC34A2, Cap1, FABP3, LALBA, SCD) were significantly down-regulated in milk purified MEC. Six were also significantly down-regulated in mammary biopsies. These transcripts are mostly involved in milk synthesis. We therefore conclude that milk purified MEC cannot be used as general markers of variations occurring in the mammary tissue but that variations in some transcripts listed above can be useful indicators. In this experimental design, udder halves were unilaterally milked once daily and the contralateral udder halves twice daily. Each mammary biopsy RNA sample from one udder half was compared to the one of the contralateral udder half in 5 dye-swaps corresponding to 5 animal replications, minimizing the animal variability. One dye swap corresponded to one comparison meaning two slides.

Project description:In a recent study, a teat disinfectant based on a nisin-producing Lactococcus cremoris displayed comparable efficacy to a conventional iodophor disinfectant in preventing dairy cow mastitis when implemented in the dairy cow milking routine. Here, we carried out a longitudinal study for an in-depth investigation of the milk proteome. Four groups of matched quarter milk with low and high somatic cell count (SCC) collected at the beginning (T0) and at the end (TF) of the trial were selected for analysis, for a total of 28 low SCC (LSCC) and 12 high SCC (HSCC) samples. The milk was delipidated, digested into peptides, and subjected to a proteomic analysis workflow entailing NanoHPLC separation and tandem mass spectrometry (LC-MS/MS) on an Orbitrap Fusion Tribrid mass spectrometer. The proteins were identified with MaxQuant, the proteomes were compared for identifying differential proteins, and their interaction networks were assessed with STRING. In LSCC milk at T0, 31 were increased or present only in the Lactococcus (L) disinfectant group (from now INCREASED in L) and 59 proteins were decreased in L disinfectant group or present only in the conventional iodophor (C) group (from now DECREASED in L). All were involved in metabolic pathways and were likely related to physiological differences between animals. In LSCC milk at TF, 34 proteins were INCREASED in L and 46 proteins were DECREASED in the L disinfectant group. Most belonged to physiological milk pathways, but 3 antimicrobial proteins were enriched in the L group at the end of the trial. In HSCC milk at T0, 54 proteins were INCREASED in L and 34 DECREASED in L. Most were involved in physiological metabolic pathways; 4 proteins with antimicrobial functions were higher in L at the beginning of the trial. In HSCC milk at TF, 67 proteins were INCREASED in L and 39 DECREASED in L. All the proteins enriched in C belonged to metabolic pathways, while up to 14 proteins enriched in L were involved in antibiotic, antimicrobial, and immune defense pathways. Cathelicidins were among the most prominent differential proteins with antimicrobial functions. At T0, cathelicidins were generally higher in HSCC than LSCC milk of both experimental groups, as expected. At TF, they were increased or unique only in the Lactococcus group, in both LSCC and HSCC milk. Western immunoblotting validation confirmed the proteomic findings. On the other hand, enzymatic assessment of n-acetyl-glucosaminidase (NAGase) did not highlight significant differences between the C and L groups, neither for LSCC nor for HSCC milk. In conclusion, the two disinfectants did not induce relevant functional changes in the milk proteome of healthy animals, except for three increased antimicrobial proteins in the Lactococcus group. On the other hand, in animals with an underlying mammary gland inflammation, the milk proteome of the Lactococcus group showed increased levels of several proteins with antibiotic, antimicrobial and immune defense functions. Among other factors, the selective immunostimulatory action of nisin, the antimicrobial effector of L. cremoris, might play a role in this observation.

Dataset Information

Gene-expression profile of the response to E. coli (k2bh2) infection in the bovine udder

Publications

In depth analysis of genes and pathways of the mammary gland involved in the pathogenesis of bovine Escherichia coli-mastitis.

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