过表达NR4A1促进山羊皮下脂肪细胞分化

doi:10.11843/j.issn.0366-6964.2021.05.012

Abstract

Abstract: The aim of this research was to clone the goat NR4A1 gene CDS sequence, clarify its tissue and cell expression patterns, and explore the effect of overexpression of NR4A1 gene on goat subcutaneous adipocyte differentiation. In this experiment, the goat overexpression vector pcDNA3.1-NR4A1 was constructed by double digestion. Five healthy Jianzhou big ear goats of one-year-old were used as experimental animals. The coding region sequence of NR4A1 gene was cloned through RT-PCR. Relative expression level of NR4A1 gene in different tissues and subcutaneous adipocytes at different stages were detected by real-time quantitative PCR (qPCR). By transfecting pcDNA3.1-NR4A1 vector into subcutaneous adipocytes to overexpress NR4A1, and changes of lipid droplet aggregation after overexpression was detected by morphology, at the same time, the relative expression level of adipose differentiation marker genes was detected by qPCR. The results showed that the coding region sequence of goat NR4A1 gene was 1 797 bp, encoding 598 amino acids. NR4A1 widely expressed in different tissues of goats and had the highest expression level in the longissimus dorsi (P<0.01), and NR4A1 expression level was the highest in goat subcutaneous adipocytes at 60 h of differentiation (P<0.01). The overexpression of NR4A1 significantly promoted the accumulation of lipid droplets in subcutaneous adipocytes of goats, and significantly upregulated the relative expression level of C/EBPα, C/EBPβ, PPARγ, LPL, SREBP1 and AP2 (P<0.05). NR4A1 maybe a positive regulator for goat subcutaneous adipocytes differentiation by regulating the expression of adipocyte differentiation marker genes.

Key words: goats, NR4A1, clone, overexpression, subcutaneous adipocyte

CLC Number:

S827.2

CUI Sheng, WANG Yong, ZHU Jiangjiang, XIONG Yan, LIN Yaqiu. Overexpression of NR4A1 Promotes Goat Subcutaneous Adipocytes Differentiation[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(5): 1258-1266.

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Overexpression of NR4A1 Promotes Goat Subcutaneous Adipocytes Differentiation

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