Zheng Yue, Chen Kun-lin, Zheng Xiao-min, Li Hui-xia, Wang Gen-lin
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.
Cell Stress Chaperones. 2014 Nov;19(6):973-81. doi: 10.1007/s12192-014-0521-8. Epub 2014 Jun 11.
MicroRNAs (miRNAs) are small single-stranded non-coding RNAs that have an important regulatory function in animal growth and developmental processes. However, the differential expression of miRNA and the role of these miRNAs in heat-stressed Holstein cows are still unknown. In this study, the profile of differentially expressed miRNAs and the target genes analysis in the serum of heat-stressed and normal Holstein cows were investigated by a Solexa deep-sequencing approach and bioinformatics. The data identified 52 differentially expressed miRNAs in 486 known miRNAs which were changed significantly between heat-stressed and normal Holstein cows (fold change >2, P < 0.001). Target genes analysis showed that at least 7 miRNAs (miR-19a, miR-19b, miR-146a, miR-30a-5p, miR-345-3p, miR-199a-3p, and miR-1246) were involved in the response to stress, oxidative stress, development of the immune system, and immune response among the identified 52 differentially expressed miRNAs. Five miRNAs (miR-27b, miR-181a, miR-181b, miR-26a, and miR-146b) were involved in stress and immune responses and the expression of five miRNAs was striking (P < 0.001). In addition, RT-qPCR and deep-sequencing methods showed that 8 miRNAs among the 12 selected miRNAs (miR-19a, miR-19b, miR-27b, miR-30a-5p, miR-181a, miR-181b, miR-345-3p, and miR-1246) were highly expressed in the serum of heat-stressed Holstein cows. GO and KEGG pathway analysis showed that these differentially expressed miRNAs were involved in a pathway that may differentially regulate the expression of stress response and immune response genes. Our study provides an overview of miRNAs expression profile and the interaction between miRNAs and their target genes, which will lead to further understanding of the important roles of miRNAs in heat-stressed Holstein cows.
微小RNA（miRNA）是一类小的单链非编码RNA，在动物生长和发育过程中具有重要的调节功能。然而，miRNA的差异表达及其在热应激荷斯坦奶牛中的作用仍不清楚。在本研究中，通过Solexa深度测序方法和生物信息学，对热应激和正常荷斯坦奶牛血清中差异表达的miRNA谱及其靶基因进行了分析。数据鉴定出在486个已知miRNA中有52个差异表达的miRNA，它们在热应激和正常荷斯坦奶牛之间有显著变化（折叠变化>2，P<0.001）。靶基因分析表明，在所鉴定的52个差异表达的miRNA中，至少有7个miRNA（miR-19a、miR-19b、miR-146a、miR-30a-5p、miR-345-3p、miR-199a-3p和miR-1246）参与应激反应、氧化应激、免疫系统发育和免疫反应。5个miRNA（miR-27b、miR-181a、miR-181b、miR-26a和miR-146b）参与应激和免疫反应，且这5个miRNA的表达显著（P<0.001）。此外，RT-qPCR和深度测序方法显示，在所选的12个miRNA（miR-19a、miR-19b、miR-27b、miR-30a-5p、miR-181a、miR-181b、miR-345-3p和miR-1246）中有8个在热应激荷斯坦奶牛血清中高表达。GO和KEGG通路分析表明，这些差异表达的miRNA参与了一个可能差异调节应激反应和免疫反应基因表达的通路。我们的研究概述了miRNA的表达谱以及miRNA与其靶基因之间的相互作用，这将有助于进一步了解miRNA在热应激荷斯坦奶牛中的重要作用。
