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Published on: January 30, 2017
Development and validation of a bovine macrophage specific cDNA microarray
Kirsty Jensen1, Richard Talbot, Edith Paxton
1Division of Genetics & Genomics, Roslin Institute, Roslin, Midlothian, Edinburgh, EH25 9PS, UK. Kirsty.Jensen@bbsrc.ac.uk
Insights
Researchers developed a bovine macrophage-specific microarray to study gene expression in cattle immune cells. This new tool, containing over 5000 genes, aids in understanding immune responses in livestock.
Area of Science:
- Immunology
- Genomics
- Livestock Science
Background:
- Macrophage response to danger signals is crucial for the early immune response.
- Microarray analysis enables large-scale gene expression studies.
- Limited microarray resources exist for livestock immunology research.
Purpose of the Study:
- To develop a bovine macrophage-specific (BoMP) cDNA microarray.
- To provide a tool for investigating gene expression in bovine myeloid cells.
Main Methods:
- Development of a BoMP cDNA microarray with 5026 sequence elements.
- Generation of a BoMP cDNA library from stimulated bovine myeloid cells.
- Validation using bovine monocytes stimulated with interferon-gamma and lipopolysaccharide.
Main Results:
- The BoMP microarray contains over 5000 unique bovine gene sequences.
- Stimulation of bovine monocytes revealed 695 differentially expressed genes.
- Key genes identified include interleukin 6, prion protein, and toll-like receptor 4.
Conclusions:
- A 5K cDNA microarray for bovine myeloid cells has been successfully created.
- The BoMP microarray is a valuable resource for livestock immunology research.
- This tool is available from the ARK-Genomics Centre.
Background:
The response of macrophages to danger signals is an important early stage in the immune response. Our understanding of this complex event has been furthered by microarray analysis, which allows the simultaneous investigation of the expression of large numbers of genes. However, the microarray resources available to study these events in livestock animals are limited.
Results:
Here we report the development of a bovine macrophage specific (BoMP) cDNA microarray. The BoMP microarray contains 5026 sequence elements (printed in duplicate) and numerous controls. The majority of the clones incorporated on the microarray were derived from the BoMP cDNA library generated from bovine myeloid cells subjected to various stimuli, including over 900 sequences unique to the library. Additional clones representing immunologically important genes have been included on the BoMP microarray. The microarray was validated by investigating the response of bovine monocytes to stimulation with interferon-gamma and lipopolysaccharide using amplified RNA. At 2 and 16 hours post stimulation 695 genes exhibited statistically significant differential expression, including; 26 sequences unique to the BoMP library, interleukin 6, prion protein and toll-like receptor 4.
Conclusion:
A 5 K cDNA microarray has been successfully developed to investigate gene expression in bovine myeloid cells. The BoMP microarray is available from the ARK-Genomics Centre for Functional Genomics in Farm Animals, UK.
