J Vet Sci.  2015 Sep;16(3):253-263. 10.4142/jvs.2015.16.3.253.

Comparative proteomic analysis of proteins expression changes in the mammary tissue of cows infected with Escherichia coli mastitis

• Zhao XW
• Yang YX
• Huang DW
• Cheng GL
• Zhao HL

• KMID: 2344297
• DOI: http://doi.org/10.4142/jvs.2015.16.3.253

Abstract

Cows infected with Escherichia (E.) coli usually experience severe clinical symptoms, including damage to mammary tissues, reduced milk yield, and altered milk composition. In order to investigate the host response to E. coli infection and discover novel markers for mastitis treatment, mammary tissue samples were collected from healthy cows and bovines with naturally occurring severe E. coli mastitis. Changes of mammary tissue proteins were examined using two-dimensional gel electrophoresis and label-free proteomic approaches. A total of 95 differentially expressed proteins were identified. Of these, 56 proteins were categorized according to molecular function, cellular component, and biological processes. The most frequent biological processes influenced by the proteins were response to stress, transport, and establishment of localization. Furthermore, a network analysis of the proteins with altered expression in mammary tissues demonstrated that these factors are predominantly involved with binding and structural molecule activities. Vimentin and alpha-enolase were central "functional hubs" in the network. Based on results from the present study, disease-induced alterations of protein expression in mammary glands and potential markers for the effective treatment of E. coli mastitis were identified. These data have also helped elucidate defense mechanisms that protect the mammary glands and promote the pathogenesis of E. coli mastitis.

Keyword

dairy cows; Escherichia coli; mammary tissue; mastitis; proteome

MeSH Terms

Animals
Cattle
Electrophoresis, Gel, Two-Dimensional/veterinary
Escherichia coli/*physiology
Escherichia coli Infections/genetics/immunology/microbiology/*veterinary
Female
Mammary Glands, Animal/*immunology/pathology
Mastitis, Bovine/*genetics/immunology/microbiology
Proteome/*genetics/metabolism
*Proteomics
Proteome

Figure

• 

  Fig. 1 Changes in the protein spots detected on two-dimensional gel electrophoresis (2-DE) gels of mammary tissues from healthy cows and animals with Escherichia (E.) coli mastitis. (A) 2-DE gel for mammary tissues from healthy cows. (B) 2-DE gel for mammary tissues from cows with severe E. coli mastitis. The labels and arrows indicating spots corresponding to proteins with altered expression levels.

• 

  Fig. 2 Abundance of β-casein in mammary tissues from cows with severe E. coli mastitis and healthy animals. (A) Representative images of spots corresponding to β-casein in the 2-DE map. (B) Densitometric analysis of the β-casein spots in the 2-DE map. (C) Western blot analysis of β-casein expression. (D) Densitometric analysis of the Western blot. Values are presented as the mean ± standard error (SE). *p < 0.05 (n = 3).

• 

  Fig. 3 Classification of differentially expressed proteins in mammary tissues from healthy cows and bovines with E. coli mastitis based on gene ontology annotations. (A) Molecular function. (B) Cellular component. (C) Biological process.

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  Fig. 4 Protein-protein network of differentially expressed proteins in mammary tissues from control cows and cows with E. coli mastitis. (A) Graphic depiction of the degree of connections for each protein in the network. (B) Representative image of the protein-protein interaction network. Pink lines indicate the connections confirmed by the experimental study, the blue lines indicate connections derived from the databases, and the yellow lines indicate connections described in abstracts of articles published in the literature.

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