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| Molecular Characteristics and Expression Analysis of amhr2 Gene in Bastard Halibut Paralichthys olivaceus |
| ZHANG Fayang1, LI Kun1, WANG Weichao1, ZHANG Dandan1, FU Xiaona1, LIU Cui1, ZHANG Junling1,2,3 |
1. Key Laboratory of Exploration and Utilization of Aquatic Germplasm Resources, Ministry of Education, Shanghai Ocean University, Shanghai 201306, China;
2. Aquatic Animal Genetic Breeding Center Shanghai Collaborative Innovation Center, Shanghai Ocean University, Shanghai 201306, China;
3. International Research Center for Marine Biosciences, Shanghai Ocean University, Shanghai 201306, China |
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Abstract In order to investigate the molecular characteristics of anti-Müllerian hormone (AMH) and its type Ⅱ receptor AMHR2 and its role in gonadal development, the amhr2 gene was cloned in bastard halibut Paralichthys olivaceus with gonads from stage Ⅲ to stage Ⅳ and body weight of (720±50) g and identified by molecular cloning and bioinformatics methods. The expression profile of amhr2 gene was determined in gonad, brain, stomach, muscle, liver, kidney and heart of bastard halibut by semi-quantitative PCR and real-time fluorescence quantitative PCR and chemical in situ hybridization (ISH). The results showed that the amhr2 cDNA sequence obtained by cloning and splicing was 1536 bp in length and encoded 511 amino acids. The amino acid sequence analysis showed that the near N-terminus of the Amhr2 protein contained a signal peptide structure of 19 amino acids (1st—19th amino acids) and the amino acid sequence comparison showed that it had the maximal similarity with Hippoglossus stenolepis (89.96%). The evolutionary tree analysis revealed that the amhr2 gene of bastard halibut was the most closely related to H. stenolepis and turbot Scophthalmus maximus, and its evolutionary status was consistent, indicating that the amhr2 gene of bastard halibut is conserved with other species. The quantitative results showed that the amhr2 gene was mainly expressed in the ovaries and testes, with significantly higher expression level in the ovaries than that in the testes. The in situ hybridization results showed that amhr2 gene was mainly expressed in the sertoli cells, primary spermatocytes and secondary spermatocytes of the testis, and expressed in the interstitial cells, Ⅱ—Ⅴ oocytes and follicular membrane in the ovary. The amhr2 gene is most closely related to the H. stenolepis and expressed primarily in gonad and in both germ cells and somatic cells of the testis and ovary, which may be important for the regulation of gonadal development in the bastard halibut.
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Received: 17 February 2023
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