鸡微粒体甘油三酯转运蛋白样基因生物学特性及表达调控

doi:10.11843/j.issn.0366-6964.2022.09.008

Abstract

Abstract: The purpose of the study was to explore the biological characteristics and expression regulation of chicken microsomal triglyceride transfer protein like gene (MTTPL), and lay a foundation for further studying its biological function in lipid metabolism of chicken liver. In this study, firstly, PCR and sequencing were used to clone MTTPL cDNA sequence. Online softwares were used to analyze the structural domain, three-dimensional structure of MTTPL and construct phylogenetic tree. Overexpression vector pcDNA3.1-MTTPL-EGFP was constructed, and co-transfected chicken hepatocellular carcinoma (LMH) with DsRed-λ vector, which expressed endoplasmic reticulum marker protein, for MTTPL subcellular localization. Then tissue samples were collected from Lushi blue-shelled chickens at different weeks of age (8 chickens for 1 day old, 1 week old, 10 week old, 30 week old, respectively). The spatial and temporal expression profiles of chicken MTTPL gene and microsomal triglyceride transporter (MTTP) gene were analyzed by real-time quantitative PCR (qPCR). Then, the effects of estrogen on the expression of MTTPL and MTTP in chicken liver were estimated, twenty hy-line brown chickens in each group were treated with 17β-estradiol at 0.5, 1 and 2 mg·kg^-1 body weight for 12 and 24 h, respectively. Lastly, the regulation mechanism of estrogen on the expression of MTTPL and MTTP were studied, 1, 50 and 100 nmol·L^-1 17β-estradiol and estrogen receptor antagonist were used to treat chicken embryo hepatocytes, respectively. The results showed that chicken MTTPL CDS region was 2 646 bp in length, encoding 881 amino acids, and sharing a common ancestor with human and chicken MTTP. MTTPL was located in endoplasmic reticulum. Chicken MTTPL and MTTP had the same functional domains as human MTTP, and the similarity deviation of three-dimensional structure with human MTTP were 0.090 and 0.064, respectively. MTTPL was highly expressed in liver and kidney tissues of chicken. MTTP was highly expressed in chicken liver, kidney and small intestine. With the growth development of chickens, the expression levels of ApoB and MTTPL in liver were significantly up-regulated (P<0.05), while the expression levels of MTTP in liver was significantly increased before 10-week-old (P<0.05), but no significant change between 10-week-old and 30-week-old chickens (P>0.05). After estrogen treatment for 12 and 24 h, the expression levels of MTTPL and ApoB in chicken liver were significantly up-regulated (P<0.05). MTTP was significantly down-regulated in high concentration group (P<0.05), while no significant change in low concentration group. Compared with the control, estrogen could significantly up-regulate the expression level of ApoB and MTTPL in chicken embryo hepatocytes (P<0.05), while MTTP was not significantly changed. Compared with estrogen treatment group, the expression level of MTTPL gene was significantly decreased in estrogen and ERα antagonist MPP co-treated group (P<0.05). Compared with MPP and ERα antagonist co-treated group, no significant change of MTTPL expression level in ERα and ERβ receptor antagonist ICI or TAM and estrogen co-treated group was observed. In conclusion, chicken MTTPL was located in endoplasmic reticulum, and had the same functional domains as human MTTP. Both MTTPL and MTTP were highly expressed in chicken liver. Compared with pre-laying stage, MTTPL expression in liver of chicken at laying stage was significantly up-regulated, while no significant difference was observed in MTTP expression. Estrogen could regulate the expression of MTTPL via binding with ERα receptor, but MTTP was not affected. It was showed that MTTPL may play an important role in liver lipid metabolism of chicken at the egg-laying stage.

Key words: chicken, MTTP, MTTPL, lipid metabolism, estrogen, lipoprotein

CLC Number:

S831.2

XU Chunlin, CAO Yuzhu, XIA Tian, JIA Qihui, WANG Dandan, ZHENG Hang, TIAN Yadong, KANG Xiangtao, JIANG Ruirui, LIU Xiaojun, LI Hong. Biological Characteristics and Expression Regulation of Chicken Microsomal Triglyceride Transfer Protein Like Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 2900-2911.

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Biological Characteristics and Expression Regulation of Chicken Microsomal Triglyceride Transfer Protein Like Gene

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