全基因组调查和功能分析揭示 TCF21 通过直接上调 HTR2A 促进鸡前体脂肪细胞分化。
Genome-wide survey and functional analysis reveal TCF21 promotes chicken preadipocyte differentiation by directly upregulating HTR2A.
机构信息
Key Laboratory of Chicken Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Harbin, 150030, Heilongjiang, China; Key Laboratory of Animal Genetics, Breeding and Reproduction, Education Department of Heilongjiang Province, Harbin, 150030, Heilongjiang, China; College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, Heilongjiang, China.
Key Laboratory of Chicken Genetics and Breeding, Ministry of Agriculture and Rural Affairs, Harbin, 150030, Heilongjiang, China; Key Laboratory of Animal Genetics, Breeding and Reproduction, Education Department of Heilongjiang Province, Harbin, 150030, Heilongjiang, China; College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, Heilongjiang, China.
出版信息
Biochem Biophys Res Commun. 2022 Jan 8;587:131-138. doi: 10.1016/j.bbrc.2021.11.103. Epub 2021 Nov 30.
BACKGROUND/AIM: Previously, we showed that transcription factor 21 (TCF21) promotes chicken preadipocyte differentiation. However, the genome-wide TCF21 binding sites and its downstream target genes in chicken adipogenesis were unknown.
METHODS
ChIP-Seq and RNA-Seq were used to screen candidate targets of TCF21. qPCR and luciferase reporter assay were applied to verify the sequencing results. Western blotting, oil red-O staining and pharmacological treatments were performed to investigate the function of 5-hydroxytryptamine receptor 2A (HTR2A), one of the bonafide direct downstream binding targets of TCF21.
RESULTS
A total of 94 candidate target genes of TCF21 were identified. ChIP-qPCR, RT-qPCR, and luciferase reporter assay demonstrated that HTR2A is one of the bonafide direct downstream binding targets of TCF21. HTR2A expression in adipose tissue was upregulated in fat line broilers. Also, the abundance of HTR2A gradually increased during the adipogenesis process. Interestingly, pharmacological enhancement or inhibition of HTR2A promoted or attenuated the differentiation of preadipocytes, respectively. Furthermore, HTR2A inhibition impaired the TCF21 promoted adipogenesis.
CONCLUSIONS
We profiled the genome-wide TCF21 binding sites in chicken differentiated preadipocytes revealing HTR2A as the direct downstream target of TCF21 in adipogenesis.
背景/目的：之前，我们表明转录因子 21（TCF21）促进鸡前体脂肪细胞分化。然而，在鸡脂肪生成过程中，TCF21 的全基因组结合位点及其下游靶基因尚不清楚。
方法
使用 ChIP-Seq 和 RNA-Seq 筛选 TCF21 的候选靶标。应用 qPCR 和荧光素酶报告基因检测来验证测序结果。通过 Western blot、油红 O 染色和药理学处理来研究 TCF21 的一个真正的直接下游结合靶标 5-羟色胺受体 2A（HTR2A）的功能。
结果
共鉴定出 94 个 TCF21 的候选靶基因。ChIP-qPCR、RT-qPCR 和荧光素酶报告基因检测表明 HTR2A 是 TCF21 的真正的直接下游结合靶标之一。在脂肪鸡中，脂肪组织中的 HTR2A 表达上调。此外，HTR2A 的丰度在脂肪生成过程中逐渐增加。有趣的是，药理学增强或抑制 HTR2A 分别促进或减弱前脂肪细胞的分化。此外，HTR2A 抑制损害了 TCF21 促进的脂肪生成。
结论
我们对鸡分化前体脂肪细胞中的 TCF21 全基因组结合位点进行了分析，揭示了 HTR2A 是脂肪生成中 TCF21 的直接下游靶标。
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