Liu Haibo, Nguyen Yet T, Nettleton Dan, Dekkers Jack C M, Tuggle Christopher K
Department of Animal Science, Iowa State University, 2258 Kildee Hall, Ames, IA, 50011, USA.
Department of Statistics, Iowa State University, 1121 Snedecor Hall, Ames, IA, 50011, USA.
BMC Genomics. 2016 Jan 22;17:73. doi: 10.1186/s12864-016-2395-x.
Improving feed efficiency (FE) of pigs by genetic selection is of economic and environmental significance. An increasingly accepted measure of feed efficiency is residual feed intake (RFI). Currently, the molecular mechanisms underlying RFI are largely unknown. Additionally, to incorporate RFI into animal breeding programs, feed intake must be recorded on individual pigs, which is costly and time-consuming. Thus, convenient and predictive biomarkers for RFI that can be measured at an early age are greatly desired. In this study, we aimed to explore whether differences exist in the global gene expression profiles of peripheral blood of 35 to 42 day-old pigs with extremely low (more efficient) and high RFI (less efficient) values from two lines that were divergently selected for RFI during the grow-finish phase, to use such information to explore the potential molecular basis of RFI differences, and to initiate development of predictive biomarkers for RFI.
We identified 1972 differentially expressed genes (DEGs) (q ≤ 0.15) between the low (n = 15) and high (n = 16) RFI groups of animals by using RNA sequencing technology. We validated 24 of 37 selected DEGs by reverse transcription-quantitative PCR (RT-qPCR) in a joint analysis of 24 (12 per line) of the 31 samples already used for RNA-seq plus 24 (12 per line) novel samples from the same contemporary group of pigs. Using an analysis of the 24 novel samples alone, only nine of the 37 selected DEGs were validated. Genes involved in small molecule biosynthetic process, antigen processing and presentation of peptide antigen via major histocompatibility complex (MHC) class I, and steroid biosynthetic process were overrepresented among DEGs that had higher expression in the low versus high RFI animals. Genes known to function in the proteasome complex or mitochondrion were also significantly enriched among genes with higher expression in the low versus high RFI animals. Alternatively, genes involved in signal transduction, bone mineralization and regulation of phosphorylation were overrepresented among DEGs with lower expression in the low versus high RFI animals. The DEGs significantly overlapped with genes associated with disease, including hyperphagia, eating disorders and mitochondrial diseases (q < 1E-05). A weighted gene co-expression network analysis (WGCNA) identified four co-expression modules that were differentially expressed between the low and high RFI groups. Genes involved in lipid metabolism, regulation of bone mineralization, cellular immunity and response to stimulus were overrepresented within the two modules that were most significantly differentially expressed between the low and high RFI groups. We also found five of the DEGs and one of the co-expression modules were significantly associated with the RFI phenotype of individual animals (q < 0.05).
The post-weaning blood transcriptome was clearly different between the low and high RFI groups. The identified DEGs suggested potential differences in mitochondrial and proteasomal activities, small molecule biosynthetic process, and signal transduction between the two RFI groups and provided potential new insights into the molecular basis of RFI in pigs, although the observed relationship between the post-weaning blood gene expression and RFI phenotype measured during the grow-finish phase was not strong. DEGs and representative genes in co-expression modules that were associated with RFI phenotype provide a preliminary list for developing predictive biomarkers for RFI in pigs.
通过基因选择提高猪的饲料效率(FE)具有经济和环境意义。残余采食量(RFI)是一种越来越被认可的饲料效率衡量指标。目前,RFI潜在的分子机制在很大程度上尚不清楚。此外,要将RFI纳入动物育种计划,必须记录每头猪的采食量,这既昂贵又耗时。因此,非常需要能够在早期阶段测量的、方便且具有预测性的RFI生物标志物。在本研究中,我们旨在探究来自两个在生长育肥阶段针对RFI进行了反向选择的品系、RFI值极低(效率更高)和极高(效率更低)的35至42日龄仔猪外周血的全基因组表达谱是否存在差异,利用这些信息探索RFI差异的潜在分子基础,并启动RFI预测性生物标志物的开发。
通过RNA测序技术,我们在低RFI组(n = 15)和高RFI组(n = 16)动物之间鉴定出1972个差异表达基因(DEG)(q≤0.15)。我们在已用于RNA测序的31个样本中的24个(每个品系12个)加上来自同一同期猪群的24个(每个品系12个)新样本的联合分析中,通过逆转录定量PCR(RT-qPCR)验证了37个选定DEG中的24个。仅对24个新样本进行分析时,37个选定DEG中仅9个得到验证。在低RFI动物中表达高于高RFI动物的DEG中,参与小分子生物合成过程、通过主要组织相容性复合体(MHC)I类进行肽抗原的抗原加工和呈递以及类固醇生物合成过程的基因占比过高。在低RFI动物中表达高于高RFI动物的基因中,已知在蛋白酶体复合物或线粒体中起作用的基因也显著富集。另外,在低RFI动物中表达低于高RFI动物的DEG中,参与信号转导、骨矿化和磷酸化调节的基因占比过高。这些DEG与包括食欲亢进、饮食失调和线粒体疾病在内的疾病相关基因显著重叠(q < 1E - 05)。加权基因共表达网络分析(WGCNA)确定了四个在低RFI组和高RFI组之间差异表达的共表达模块。在低RFI组和高RFI组之间差异最显著的两个模块中,参与脂质代谢、骨矿化调节、细胞免疫和对刺激反应的基因占比过高。我们还发现5个DEG和1个共表达模块与个体动物的RFI表型显著相关(q < 0.05)。
低RFI组和高RFI组断奶后血液转录组明显不同。鉴定出的DEG表明两个RFI组在线粒体和蛋白酶体活性、小分子生物合成过程以及信号转导方面存在潜在差异,并为猪RFI的分子基础提供了潜在的新见解,尽管断奶后血液基因表达与生长育肥阶段测量的RFI表型之间观察到的关系并不强烈。与RFI表型相关的DEG和共表达模块中的代表性基因提供了开发猪RFI预测性生物标志物的初步清单。