Ahanda Marie-Laure Endale, Zerjal Tatiana, Dhorne-Pollet Sophie, Rau Andrea, Cooksey Amanda, Giuffra Elisabetta
INRA, UMR 1313 Génétique Animale et Biologie Intégrative, Jouy-en-Josas, France; CEA, DSV, IRCM, SREIT, Laboratoire de Radiobiologie et Etude du Génome, Jouy-en-Josas, France; AgroParisTech, UMR 1313 Génétique Animale et Biologie Intégrative, Paris, France.
INRA, UMR 1313 Génétique Animale et Biologie Intégrative, Jouy-en-Josas, France; AgroParisTech, UMR 1313 Génétique Animale et Biologie Intégrative, Paris, France.
PLoS One. 2014 Dec 4;9(12):e114598. doi: 10.1371/journal.pone.0114598. eCollection 2014.
Circulating extra-cellular microRNAs (miRNAs) have emerged as promising minimally invasive markers in human medicine. We evaluated miRNAs isolated from total plasma as biomarker candidates of a response to an abiotic stress (feed deprivation) in a livestock species. Two chicken lines selected for high (R+) and low (R-) residual feed intake were chosen as an experimental model because of their extreme divergence in feed intake and energy metabolism. Adult R+ and R- cocks were sampled after 16 hours of feed deprivation and again four hours after re-feeding. More than 292 million sequence reads were generated by small RNA-seq of total plasma RNA. A total of 649 mature miRNAs were identified; after quality filtering, 148 miRNAs were retained for further analyses. We identified 23 and 19 differentially abundant miRNAs between feeding conditions and between lines respectively, with only two miRNAs identified in both comparisons. We validated a panel of six differentially abundant miRNAs by RT-qPCR on a larger number of plasma samples and checked their response to feed deprivation in liver. Finally, we evaluated the conservation and tissue distribution of differentially abundant miRNAs in plasma across a variety of red jungle fowl tissues. We show that the chicken plasma miRNome reacts promptly to the alteration of the animal physiological condition driven by a feed deprivation stress. The plasma content of stress-responsive miRNAs is strongly influenced by the genetic background, with differences reflecting the phenotypic divergence acquired through long-term selection, as evidenced by the profiles of conserved miRNAs with a regulatory role in energy metabolism (gga-miR-204, gga-miR-let-7f-5p and gga-miR-122-5p). These results reinforce the emerging view in human medicine that even small genetic differences can have a considerable impact on the resolution of biomarker studies, and provide support for the emerging interest in miRNAs as potential novel and minimally invasive biomarkers for livestock species.
循环细胞外微小RNA(miRNA)已成为人类医学中颇具前景的微创标志物。我们评估了从全血浆中分离出的miRNA,将其作为家畜对非生物应激(饲料剥夺)反应的生物标志物候选物。选择两个因剩余采食量高(R +)和低(R -)而被选育的鸡品系作为实验模型,因为它们在采食量和能量代谢方面存在极大差异。成年R +和R -公鸡在饲料剥夺16小时后采样,并在重新喂食4小时后再次采样。通过对全血浆RNA进行小RNA测序,产生了超过2.92亿个序列读数。共鉴定出649个成熟miRNA;经过质量筛选后,保留了148个miRNA用于进一步分析。我们分别在喂食条件之间和品系之间鉴定出23个和19个差异丰度的miRNA,在两个比较中仅鉴定出两个miRNA。我们通过RT-qPCR在大量血浆样本上验证了一组6个差异丰度的miRNA,并检查了它们在肝脏中对饲料剥夺的反应。最后,我们评估了差异丰度的miRNA在血浆中的保守性以及在各种红原鸡组织中的组织分布。我们表明,鸡血浆miRNome对由饲料剥夺应激驱动的动物生理状况改变有迅速反应。应激反应性miRNA的血浆含量受到遗传背景的强烈影响,差异反映了通过长期选择获得的表型差异,在能量代谢中起调节作用的保守miRNA(gga-miR-204、gga-miR-let-7f-5p和gga-miR-122-5p)的谱图证明了这一点。这些结果强化了人类医学中新兴的观点,即即使是微小的基因差异也可能对生物标志物研究的分辨率产生相当大的影响,并为将miRNA作为家畜潜在的新型微创生物标志物的新兴兴趣提供了支持。