Contribution of Ielovl5a/I to Docosahexaenoic Acid Synthesis at the Transcriptional Regulation Level in Common Carp, ICyprinus carpio/I

• Published in: Animals (Basel) 2024-02, Vol.14 (4)
• Main Authors: Zhang, Hanyuan, Li, Peizhen, Zhu, Youxiu, Jiang, Yanliang, Feng, Jianxin, Zhao, Zixia, Xu, Jian
• Format: Article
• Language: English
• Subjects: Amino acids; Fresh water; Genes; Genetic transcription; Luciferase; Omega-3 fatty acids; Physiological aspects; Unsaturated fatty acids
• ISSN: 2076-2615; 2076-2615
• DOI: 10.3390/ani14040544

The expression of the elongase of the very-long-chain fatty acid 5 (elovl5) gene was investigated in this study using different molecular validation strategies. It has been found that the elovl5a gene is a causal gene responsible for DHA content differences in common carp. miR-26a-5P is a casual miRNA that negatively regulates DHA synthesis by targeting the elovl5a gene. Considering its down-regulation of the DHA synthesis pathways in common carp, knocking out, knocking down, or silencing the expression of elovl5a may increase DHA content in common carp. Overall, our study preliminarily explored the potential function of elovl5a in common carp. These results provide useful information for future functional research of the elovl5 gene and the interactions between its other gene family members in freshwater fish. Docosahexaenoic acid (DHA) is an essential nutrient for humans and plays a critical role in human development and health. Freshwater fish, such as the common carp (Cyprinus carpio), have a certain degree of DHA biosynthesis ability and could be a supplemental source of human DHA needs. The elongase of very-long-chain fatty acid 5 (Elovl5) is an important enzyme affecting polyunsaturated fatty acid (PUFA) biosynthesis. However, the function and regulatory mechanism of the elovl5 gene related to DHA synthesis in freshwater fish is not clear yet. Previous studies have found that there are two copies of the elovl5 gene, elovl5a and elovl5b, which have different functions. Our research group found significant DHA content differences among individuals in Yellow River carp (Cyprinus carpio var.), and four candidate genes were found to be related to DHA synthesis through screening. In this study, the expression level of elovl5a is decreased in the high-DHA group compared to the low-DHA group, which indicated the down-regulation of elovl5a in the DHA synthesis pathways of Yellow River carp. In addition, using a dual-luciferase reporter gene assay, we found that by targeting the 3’UTR region of elovl5a, miR-26a-5p could regulate DHA synthesis in common carp. After CRISPR/Cas9 disruption of elovl5a, the DHA content in the disrupted group was significantly higher than in the wildtype group; meanwhile, the expression level of elovl5a in the disrupted group was significantly reduced compared with the wildtype group. These results suggest that elovl5a may be down-regulating DHA synthesis in Yellow River carp. This study could provide useful information for future