Using RNA-Seq to Identify Reference Genes of the Transition from Brown to White Adipose Tissue in Goats

文献类型： 外文期刊

第一作者： Wang, Linjie

作者： Wang, Linjie;Chen, Xingyue;Zhang, Xujia;Zhan, Siyuan;Cao, Jiaxue;Zhong, Tao;Guo, Jiazhong;Li, Li;Zhang, Hongping;Wang, Yan;Song, Tianzeng

作者机构：

关键词： goat; brown adipose tissue; reference gene; Normfinder; BestKeeper; geNorm

期刊名称：ANIMALS （ 影响因子：2.752； 五年影响因子：2.942 ）

ISSN： 2076-2615

年卷期： 2020 年 10 卷 9 期

页码：

收录情况： SCI

摘要： Simple Summary Brown adipose tissue (BAT) plays important roles in unique non-shivering thermogenesis. It is necessary to select reference genes during the transition process from brown (BAT) to white adipose tissue (WAT) for Quantitative PCR (qPCR) analysis. In this study,CTNNB,PFDN5andEIF3M, selected from RNA sequencing data, were the most suitable reference genes. The present study provides a detailed analysis of the expression stability of reference genes for the study of gene expression profiling during the transition process from BAT to WAT. Brown adipose tissues have unique non-shivering thermogenesis functions, can be found in newborn ruminate animals, and then are gradually replaced by white adipose tissues in adulthood. For the purpose of exploring the intrinsic mechanism underlying the conversion process from brown (BAT) to white adipose tissue (WAT), it is necessary to utilize Quantitative PCR (qPCR) to study gene expression profiling. In this study, we identified reference genes that were consistently expressed during the transformation from goat BAT to WAT using RNA-seq data. Then, twelve genes were evaluated as candidate reference genes for qPCR in goat perirenal adipose tissue using three tools (geNorm, Normfinder, and BestKeeper). In addition, the selected reference genes were used to normalize the gene expression ofPGC-1 alpha andGPAT4. It was found that traditional reference genes, such asGAPDH,RPLP0,HPRT1, andPPIAwere not suitable for target gene normalization. In contrast,CTNNB,PFDN5, andEIF3M, selected from RNA sequencing data, showed the least variation and were recommended as the best reference genes during the transformation from BAT to WAT.

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