饲料不同脂肪源对斑马鱼elovl5、elovl2和fads2在高度不饱和脂肪酸合成中的作用影响
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华中农业大学水产学院

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国家自然科学基金面上项目(31872579)


The effects of dietary different lipid sources on elovl5, elovl2 and fads2 in highly unsaturated fatty acids biosynthesis in zebrafish
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College of Fisheries,Huazhong Agriculture University

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    摘要:

    为探究延长酶5(elovl5)、延长酶2(elovl2)和去饱和酶2(fads2)在淡水鱼高度不饱和脂肪酸(HUFA)合成中的相互作用,本实验利用CRISPR/Cas9技术,成功构建出斑马鱼elovl5-/-(以下简称E5-/-组),elovl2-/-*elovl5-/-(以下简称E2-/-*E5-/-组),elovl5-/-*fads2-/-(以下简称E5-/-*F2-/-组),elovl2-/-*elovl5-/-*fads2-/-(以下简称E2-/-*E5-/-*F2-/-组)四种突变体模型,并对四种突变体进行了大豆油饲料(SO)和亚麻芥油饲料(CO)的投喂实验,研究了斑马鱼elovl5、elovl2和fads2在HUFA合成过程中对饲料不同脂肪源的响应。结果显示,在E5-/-组和E2-/-*E5-/-组中,C18PUFA未出现明显累积,而E5-/-*F2-/-组和E2-/-*E5-/-*F2-/-组中的C18:3n-3(ALA)和C18:2n-6(LNA)呈现显著的累积。elovl2,elovl5和fads2缺失后,肝脏elovl4a和elovl4b表达明显上升。不同脂肪源投喂实验结果显示,SO饲料投喂后的E5-/-组DHA含量与WT无显著差异,E5-/-*F2-/-组较WT组DHA含量显著降低。然而,CO饲料投喂后的E5-/-组DHA含量较WT组呈现显著升高,E5-/-*F2-/-组与WT无显著差异。fads2缺失斑马鱼去饱和作用受到显著影响,表明fads2作为HUFA合成过程中的必需酶。而elovl2和elovl5缺失的表型可能会被其它HUFA合成途径所弥补。本实验证明了elovl2和elovl5在HUFA合成中存在基因间相互作用,fads2作为去饱和酶在HUFA合成中必不可少。本研究为提高淡水鱼类HUFA合成能力提供了理论基础。

    Abstract:

    In order to further explore the interaction of elongase 5 (elovl5), elongase 2 (elovl2) and desaturase 2 (fads2) in the synthesis of highly unsaturated fatty acids (HUFA) in freshwater fish, our study used CRISPR/Cas9 technology to successfully construct zebrafish elovl5-/- (hereinafter referred to as E5-/- group), elovl2-/-*elovl5-/- (hereinafter referred to as E2-/-*E5-/- group), elovl5-/-*fads2-/- (hereinafter referred to as E5-/-*F2-/- group) and elovl2-/-*elovl5-/-*fads2-/- (hereinafter referred to as E2-/-*E5-/-*F2-/- group) four mutations Soybean oil (SO) and Canola oil (CO) feeding experiments were carried out on four mutants. The responses of zebrafish elovl5, elovl2 and fads2 to different fat sources in the feed during the HUFA synthesis process were studied. The results show that in the E5-/- group and E2-/-*E5-/- group, the extension of C18PUFA to C20PUFA still exists, while the E5-/-*F2-/- group and E2-/-*E5-/-*F2-/- group C18:3n-3 (ALA) and C18:2n-6 (LNA) showed significant accumulation. After mutant of elovl2, elovl5 and fads2, elovl4a and elovl4b were induced to express. The results of fatty acid analysis showed that the DHA content of E5-/- group after SO feed was not significantly different from that of WT, and the DHA content of E5-/-*F2-/- group was significantly lower than that of WT group. However, E5-/- group after CO feed was fed The DHA content of the was significantly higher than that of the WT group, and there was no significant difference between the E5-/-*F2-/- group and the WT group. Zebrafish desaturation without fads2 is significantly affected, indicating that fads2 is an essential enzyme in the process of HUFA synthesis. The missing phenotype of elovl2 and elovl5 may be compensated by other HUFA synthesis pathways. This experiment further verified that elovl2 and elovl5 have gene interactions in HUFA synthesis, fads2 as a desaturase is essential for HUFA synthesis. This study provides a theoretical basis for improving the HUFA synthesis ability of freshwater fish.

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  • 收稿日期:2021-05-20
  • 最后修改日期:2021-07-01
  • 录用日期:2021-07-29
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