Preliminary functional study of foxl2 in Portunus trituberculatus and analysis of its related miRNA
Full
- 1. Jiangsu Provincial Key Laboratory of Marine Biotechnology, Jiangsu Ocean University, Lianyungang 222005, China
- 2. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fisheries Science, Qingdao 266071, China
- 3. Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China
- 4. Key Laboratory of Aquatic Genomics, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
The foxl2 plays essential roles in regulating ovarian differentiation, development and functional maintenance of vertebrate. However, its function in ovarian development of the swimming crab Portunus trituberculatus remains unknown. In the present study, we firstly cloned the full-length cDNA of P. trituberculatus foxl2 (Ptfoxl2) which contained a 5′ untranslated region (UTR) of 701 bp, a 3′ UTR of 211 bp, and an open reading frame (ORF) 1590 bp. Gene expression analysis in different tissues showed that Ptfoxl2 exhibited the highest expression in ovary. There were significant differences in its expression in different stages of ovarian development, and its expression was the highest in stage V. After eyestalk ablation, Ptfoxl2 expression decreased significantly. In addition, RNAi of Ptfoxl2 resulted in a significant reduction in vitellogenin expression of ovary. Taken together, these results indicated that Ptfoxl2 played an important role in the regulation of ovarian development of P. trituberculatus by inhibiting endogenous vitellogenesis in maturation stage. In order to further analyze the post-transcriptional regulation of Ptfoxl2, the miRNAs targeting Ptfoxl2 was predicted by bioinformatics analysis and the interaction between the miRNAs and Ptfoxl2 was verified using the dual luciferase reporter assay. The results showed that the relative luciferase activity was significantly reduced after co-transfection of miR-9 mimics and pmirGLO-Ptfoxl2-3′ UTR. Expression analysis showed that miR-9 exhibited opposite pattern at ovarian maturation stage and after eyestalk ablation. Collectively, the results demonstrated that miR-9 can regulate Ptfoxl2 expression in the swimming crab.
Mengqian Zhang, Jingyan Zhang, Hongxing Ge, Ping Liu, Jian Li, Xianliang Meng.
Preliminary functional study of foxl2 in Portunus trituberculatus and analysis of its related miRNA[J].
Haiyang Xuebao,
2022
, 44
(4)
: 85
-94
.
DOI: 10.12284/hyxb2022060