Acta Veterinaria et Zootechnica Sinica ›› 2023, Vol. 54 ›› Issue (5): 1927-1938.doi: 10.11843/j.issn.0366-6964.2023.05.015

• ANIMAL GENETICS AND BREEDING • Previous Articles     Next Articles

APOA4 Gene Inhibits Intramuscular Adipocyte Differentiation in Goat

QUBI Wuqie1,2, LI Yanyan1,2, LI Xin1,2, WANG Yong1,2, WANG Youli1,2, LIU Wei1,2*, ZHU Jiangjiang1,2, LIN Yaqiu1,2*   

  1. 1. Key Laboratory of Protection and Utilization of Animal Genetic Resources on Qinghai-Tibet Plateau of Ministry of Education/Sichuan Province, Southwest Minzu University, Chengdu 610041, China;
    2. College of Animal Science and Veterinary Medicine, Southwest Minzu University, Chengdu 610041, China
  • Received:2022-09-09 Online:2023-05-23 Published:2023-05-20

Abstract: The study aimed to determine the regulatory effect of goat apolipoprotein A-IV (APOA4) on intramuscular adipocyte differentiation. The animals used in this experiment were 1 year old Jianzhou big ear goat rams (n=5) with good physiological state. The APOA4 gene sequence of goat was cloned by RT-PCR, and the cloned sequence was analyzed by bioinformatics analysis, and the sequence characteristics were obtained. Temporal expression profiles of APOA4 in tissues and cells of goat were constructed by qPCR. By constructing the goat APOA4 gene overexpression vector, this experiment was divided into an overexpression group (pEGFP-N1- APOA4) and a negative control group (pEGFP-N1), with 3 replicates in each group and 2 samples in each replicate. Transfected pEGFP-N1-APOA4 overexpression vector and pEGFP-N1 into goat intramuscular precursor adipocytes and induced differentiation. Oil red O staining, Bodipy staining, DAPI staining, absorbance (OD490 nm) and qPCR were used to determine the effects of overexpression of goat APOA4 gene on adipocyte differentiation and adipogenesis markers. The results showed as follows:1) The APOA4 gene sequence of goat was 1 467 bp, in which the CDS region was 1 143 bp, encoding 380 AA. 2) Bioinformatics analysis predicted that APOA4 was negatively charged and acid-unstable hydrophobic protein. 3) APOA4 was widely expressed in goat tissues, and the highest expression level was found in liver, which was significantly higher than that in other tissues (P<0.01). APOA4 expression was the highest in intramuscular adipocytes at 48 h after induction of differentiation. 4) Compared with the control group after APOA4 gene overexpression, the accumulation of lipid droplets in intramuscular adipocytes was decreased, and the expression levels of adipose-differentiation markers C/EBPα and PPARγ were significantly down-regulated (P< 0.01), and the expression levels of SREBP1 and C/EBPβ were significantly down-regulated (P<0.05). Lipid metabolism marker genes AP2 and LPL were significantly down-regulated (P<0.05). Based on morphological staining and molecular biology results, APOA4 gene may inhibit the differentiation of adipocytes and lipid droplet accumulation in goats by inhibiting the expression of differentiation markers C/EBPα, PPARγ, SREBP1, C/EBPβ and lipid metabolism markers AP2 and LPL. It was suggested that overexpression of APOA4 gene inhibited the differentiation of intramuscular adipocytes.

Key words: goat, APOA4, intramuscular adipocytes, overexpression, cell differentiation

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