Genome-Wide Identification and Functional Prediction of Circular RNAs in Response to Heat Stress in Chinese Holstein Cows

Abstract Background: Heat stress (HS) leads to substantial economic loss of dairy industry each year. The negative effect of HS in dairy cows is becoming one of the more urgent and important issue due to accelerating side-effects of global warming. At present, the best way to alleviate the effect of HS is breeding of thermotolerant cows which depends on the profound understanding of mechanism of HS. Various genes are involved in HS response but the information about the role of noncoding RNAs, especially circular RNAs (circRNAs) is largely unknown.Results: In our study, we aimed to investigate the different expression profile of circRNAs between HS and normal condition (NC) of Chinese Holstein cow’s mammary gland. CircRNAs were identified using RNA sequencing and bioinformatics analysis. In total, 37405 circRNAs were detected and 95 were differentially expressed (DE), including 15 downregulated and 80 upregulated circRNAs in HS group compared to NC. Eight circRNAs were randomly selected to verify the RNA sequencing result. Further, Sanger sequencing validated the backsplicing site of the eight circRNAs. Moreover, results obtained from the Quantitative real time PCR (qRT-PCR) showed consistent expression trend with that of RNA sequencing. GO annotation and KEGG analysis suggested that these DE circRNAs probably involved in the energy metabolic regulation. Furthermore, we constructed ceRNA network and the result indicated that these DE circRNAs could regulate lactation through IGF1 and PRL signaling pathway.Conclusion: During our study, it was evident that circRNAs were involved in regulation of HS response in Chinese Holstein cow’s mammary gland. This will help researchers to achieve a better understanding of mechanism related to HS response and can provide new insights in breeding of thermotolerant dairy cows.