Abstract
BackgroundMicroarray transcript profiling has the potential to illuminate the molecular processes that are involved in the responses of cattle to disease challenges. This knowledge may allow the development of strategies that exploit these genes to enhance resistance to disease in an individual or animal population.ResultsThe Bovine Innate Immune Microarray developed in this study consists of 1480 characterised genes identified by literature searches, 31 positive and negative control elements and 5376 cDNAs derived from subtracted and normalised libraries. The cDNA libraries were produced from 'challenged' bovine epithelial and leukocyte cells. The microarray was found to have a limit of detection of 1 pg/μg of total RNA and a mean slide-to-slide correlation co-efficient of 0.88. The profiles of differentially expressed genes from Concanavalin A (ConA) stimulated bovine peripheral blood lymphocytes were determined. Three distinct profiles highlighted 19 genes that were rapidly up-regulated within 30 minutes and returned to basal levels by 24 h; 76 genes that were up-regulated between 2–8 hours and sustained high levels of expression until 24 h and 10 genes that were down-regulated. Quantitative real-time RT-PCR on selected genes was used to confirm the results from the microarray analysis. The results indicate that there is a dynamic process involving gene activation and regulatory mechanisms re-establishing homeostasis in the ConA activated lymphocytes. The Bovine Innate Immune Microarray was also used to determine the cross-species hybridisation capabilities of an ovine PBL sample.ConclusionThe Bovine Innate Immune Microarray has been developed which contains a set of well-characterised genes and anonymous cDNAs from a number of different bovine cell types. The microarray can be used to determine the gene expression profiles underlying innate immune responses in cattle and sheep.
Highlights
Microarray transcript profiling has the potential to illuminate the molecular processes that are involved in the responses of cattle to disease challenges
In this study "signal" refers to the background subtracted mean fluorescence intensity and "element" refers to the DNA amplicon printed onto the microarray slide surface
Plasmid clones corresponding to the majority of the candidate genes were identified in available cDNA libraries including; Meat Animal Research Center (MARC) 1–5 libraries [25,26]. and CSIRO Livestock Industries ovine and bovine cDNA libraries [27,28]
Summary
Microarray transcript profiling has the potential to illuminate the molecular processes that are involved in the responses of cattle to disease challenges This knowledge may allow the development of strategies that exploit these genes to enhance resistance to disease in an individual or animal population. Specialised or focused bovine cDNA microarrays have been reported, which are suitable for studies with specific tissues or physiological states These microarrays provide an excellent tool for examination of gene expression in a specific tissue There are reports of a relatively small bovine immune-endocrine cDNA microarray representing 167 genes [4] and a third generation immune gene cDNA microarray constructed from bovine leukocytes which contains 1250 genes [10] Both of these microarrays contain only a limited representation of the many immune related genes, based on surveys of the murine and human scientific literature. A bovine Affymetrix microarray has been released the corresponding gene annotations are limited and the technology is still relatively expensive [12]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
