Characterization and Expression Analyses of Prostaglandin E2 Receptor EP4 Gene in Procambarus clarkii
JIANG Hongxia, ZHANG Ran, LI Yizheng, LIU Xuewei, ZHANG Shuaishuai, YU Miao, WANG Lei, ZHANG Meng, QIAO Zhigang, LI Xuejun
Engineering Technology Research Center of Henan Province for Aquatic Animal Cultivation, Engineering Laboratory of Henan Province for Aquatic Animal Disease Control, College of Fisheries, Henan Normal University,Xinxiang 453007, China
Abstract：Prostaglandin E2 (PGE2) receptor EP4 plays an important role in the regulation of ovarian development, ovulation and reproduction in animals. The cDNA sequence of the prostaglandin E2 (PGE2) receptor EP4 gene with the complete ORF region was obtained from the ovary of Procambarus clarkii using transcriptome sequencing. In order to explore the sequence characteristics and the function of EP4 gene in ovary of P. clarkii (PcEP4), the gene sequence was analyzed by bioinformatics, and the gene expression pattern was detected by real-time fluorescent quantitative PCR. Bioinformatics analysis results showed that the ORF of PcEP4 was 2505 bp, encoding a deduced protein with 835 amino acids; PcEP4 is a typical G protein coupled receptor with 7 transmembrane α-helical structures; EP4 amino acid sequence of P. clarkii and Litopenaeus vannamei were the most consistent,and the genetic relationship was the closest. Gene expression analysis results showed that PcEP4 gene expression level was the highest in the ovary in P. clarkii; in the Ⅰ—Ⅵ stage of ovaries in P. clarkii, PcEP4 gene expression level was the highest in stage Ⅴ ovary; At 36, 48 and 60 h after PGE2 injection, the expression level of PcEP4 gene in the ovaries of P. clarkii was significantly higher than that in the control group (P<0.05), the egg holding rate of P. clarkii also increased compared with the control group at 72 h after PGE2 injection. The above results indicate that, as a PGE2 receptor, EP4 plays an important role in promoting oocyte maturation and ovulation of P. clarkii together with PGE2.
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2023, Vol. 42 
