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| Status |
Public on Feb 28, 2012 |
| Title |
Alteration of gene expression in mouse mammary tissue upon E. coli infection |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Mastitis is a multietiological complex disease, which is defined as inflammation of parenchyma of mammary glands. Bacterial infection is the predominant cause of mastitis, though fungal, viral and mycoplasma infections also have been reported. Based on severity of the disease, mastitis can be classified into- subclinical, clinical and chronic forms. Bacterial pathogens from the fresh cow milk were isolated and classified by standard microbiological tests and multiplex PCR. Epidemiological studies have shown that Escherichia coli is the 2nd largest mastitic pathogen after Staphylococcus aureus in India. Based on Enterobacterial Repetitive Intergenic Consensus (ERIC) -PCR profile and presence of virulence genes, a field isolate of E. coli was used for intramammary inoculation in lactating mice. Histopathological examination of H&E stained sections showed severe infiltration of polymorphonuclear neutrophils (PMNs), mononuclear inflammatory cells in the alveolar lumen and also in interstitial space and necrosis of alveolar epithelial cells after 24h. Western blot and immunohistochemical analysis of the mice mammary tissues have shown significant hyperacetylation at histone H3Lys14 residue of both mammary epithelial cells and migrated inflammatory cells. Quantitative real-time PCR and genome-wide gene expression profile in the E. coli infected mice mammary tissue revealed differential expression of genes related to inflammation, immunity, antimicrobial peptide expression, acute phase response and oxidative stress response. Expression levels of milk proteins were also suppressed. ChIP assay from the paraffinized tissues have shown selective enrichment of acetylated histone H3K14 and H4K8 at the promoters of over expressed genes. These data suggests that E. coli infection in the mice mammary tissue leads to histone hyperacetylation at the immune gene promoters which is pre-requisite for expression of inflammatory genes to mount drastic immune response.
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| Overall design |
Abdominal mammary glands of 7 Day post partum lactating female Swiss albino mice was challenged with a pure culture of E. coli isolated from cow milk. Mammary tissues were harvested after 24h of infection. Total RNA was isolated by Qiagen RNeasy mini columns and used for global gene expression and RT-PCR analysis. For RT-PCR, cDNAs corresponding to total mRNAs were synthesized using Oligo-dT primer. Global gene expression analysis were done by single colour labelling of total RNA and hybridization to Agilent 8x60k mouse Microarray.
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| Contributor(s) |
Modak R, Kundu TK, Shome BR, Das Mitra S |
| Citation(s) |
22419123 |
| |
| Submission date |
Feb 01, 2012 |
| Last update date |
Jan 18, 2013 |
| Contact name |
Tapas Kumar Kundu |
| E-mail(s) |
[email protected]
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| Phone |
+91-80-22082840
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| Organization name |
Jawaharlal Nehru Centre for Advanced Scientific Research
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| Department |
Molecular Biology & Genetics Unit
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| Lab |
Transcription & Disease Laboratory
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| Street address |
Jakkur P.O.
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| City |
Bangalore |
| State/province |
Karnataka |
| ZIP/Postal code |
560064 |
| Country |
India |
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| Platforms (1) |
| GPL13381 |
Custom Whole Genome Mouse 8x60k designed by Genotypic Technology Pvt. Ltd. (Agilent-026986) |
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| Samples (4)
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| Relations |
| BioProject |
PRJNA152353 |
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