Transcriptomic analysis of the stress response to weaning at housing in bovine leukocytes using RNA-seq technology.
BACKGROUND
Weaning of beef calves is a necessary husbandry practice and involves separating the calf from its mother, resulting in numerous stressful events including dietary change, social reorganisation and the cessation of the maternal-offspring bond and is often accompanied by housing. While much recent research has focused on the physiological response of the bovine immune system to stress in recent years, little is known about the molecular mechanisms modulating the immune response. Therefore, the objective of this study was to provide new insights into the molecular mechanisms underlying the physiological response to weaning at housing in beef calves using Illumina RNA-seq.


RESULTS
The leukocyte transcriptome was significantly altered for at least 7 days following either housing or weaning at housing. Analysis of differentially expressed genes revealed that four main pathways, cytokine signalling, transmembrane transport, haemostasis and G-protein-coupled receptor (GPRC) signalling were differentially regulated between control and weaned calves and underwent significant transcriptomic alterations in response to weaning stress on day 1, 2 and 7. Of particular note, chemokines, cytokines and integrins were consistently found to be up-regulated on each day following weaning. Evidence for alternative splicing of genes was also detected, indicating a number of genes involved in the innate and adaptive immune response may be alternatively transcribed, including those responsible for toll receptor cascades and T cell receptor signalling.


CONCLUSIONS
This study represents the first application of RNA-Seq technology for genomic studies in bovine leukocytes in response to weaning stress. Weaning stress induces the activation of a number of cytokine, chemokine and integrin transcripts and may alter the immune system whereby the ability of a number of cells of the innate and adaptive immune system to locate and destroy pathogens is transcriptionally enhanced. Stress alters the homeostasis of the transcriptomic environment of leukocytes for at least 7 days following weaning, indicating long term effects of stress exposure in the bovine. The identification of gene signature networks that are stress activated provides a mechanistic framework to characterise the multifaceted nature of weaning stress adaptation in beef calves. Thus, capturing subtle transcriptomic changes provides insight into the molecular mechanisms that underlie the physiological response to weaning stress.
Animal and Bioscience Research Department, Animal & Grassland Research and Innovation Centre, Grange, Dunsany, Co. Meath, Ireland.
Aran
A
O'Loughlin
David J
DJ
Lynn
Mark
M
McGee
Sean
S
Doyle
Matthew
M
McCabe
Bernadette
B
Earley
D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D001665:Binding Sites; D001792:Blood Platelets / Q000378:metabolism; D001831:Body Temperature / Q000235:genetics; D002417:Cattle; D019295:Computational Biology; D016207:Cytokines / Q000378:metabolism; D005091:Exons / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D006242:Haptoglobins / Q000378:metabolism; D006799:Housing, Animal; D006854:Hydrocortisone / Q000097:blood; D007962:Leukocytes / Q000378:metabolism*; D008214:Lymphocytes / Q000378:metabolism; D008297:Male; D009504:Neutrophils / Q000378:metabolism; D012313:RNA / Q000235:genetics* / Q000378:metabolism; D017423:Sequence Analysis, RNA / Q000379:methods*; D015398:Signal Transduction / Q000235:genetics; D013312:Stress, Physiological / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism; D059467:Transcriptome / Q000235:genetics; D014886:Weaning
D016207:Cytokines; D006242:Haptoglobins; D014157:Transcription Factors; D012313:RNA; D006854:Hydrocortisone