Differential neutrophil gene expression in early bovine pregnancy

Reproductive Biology and Endocrinology

Research

Ayumi Shichijo-Kizaki¹  Kazuyoshi Hashizume¹  Keiichiro Kizaki¹  Misa Hosoe²  Toru Takahashi²  Tadashi Furusawa² 

[1] Department of Veterinary Medicine, Laboratory of Veterinary Physiology,Iwate University, Ueda 3-18-8, 020-8550, Morioka, Iwate, Japan;Reproductive Biology Unit, National Institute of Agrobiological Sciences, Ikenodai 2, 305-8602, Tsukuba, Japan;

关键词: Pregnancy diagnosis;    Neutrophils;    Bovine;    ISGs;   

DOI  :  10.1186/1477-7827-11-6

 received in 2012-10-13, accepted in 2013-02-03,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundIn food production animals, especially cattle, the diagnosis of gestation is important because the timing of gestation directly affects the running of farms. Various methods have been used to detect gestation, but none of them are ideal because of problems with the timing of detection or the accuracy, simplicity, or cost of the method. A new method for detecting gestation, which involves assessing interferon-tau (IFNT)-stimulated gene expression in peripheral blood leukocytes (PBL), was recently proposed. PBL fractionation methods were used to examine whether the expression profiles of various PBL populations could be used as reliable diagnostic markers of bovine gestation.MethodsPBL were collected on days 0 (just before artificial insemination), 7, 14, 17, 21, and 28 of gestation. The gene expression levels of the PBL were assessed with microarray analysis and/or quantitative real-time reverse transcription (q) PCR. PBL fractions were collected by flow cytometry or density gradient cell separation using Histopaque 1083 or Ficoll-Conray solutions. The expression levels of four IFNT-stimulated genes, interferon-stimulated protein 15 kDa (ISG15), myxovirus-resistance (MX) 1 and 2, and 2′-5′-oligoadenylate synthetase (OAS1), were then analyzed in each fraction through day 28 of gestation using qPCR.ResultsMicroarray analysis detected 72 and 28 genes in whole PBL that were significantly higher on days 14 and 21 of gestation, respectively, than on day 0. The upregulated genes included IFNT-stimulated genes. The expression levels of these genes increased with the progression of gestation until day 21. In flow cytometry experiments, on day 14 the expression levels of all of the genes were significantly higher in the granulocyte fraction than in the other fractions. Their expression gradually decreased through day 28 of gestation. Strong correlations were observed between the expression levels of the four genes in the granulocyte fractions obtained with flow cytometry and with density gradient separation.ConclusionsThe expression profiles of ISG15, MX1, MX2, and OAS1 could be a useful diagnostic biomarker of bovine gestation. Assessing the expression levels of these genes in a granulocyte fraction obtained with density gradient separation is a practical way of detecting gestation in cows within three weeks of insemination.

【 授权许可】

Unknown   
© Kizaki et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.