Gene cloning of anti-Müllerian hormone and analysis of its mRNA level in Scatophagus argus at the different development stages of gonad

To study the role of AMH in gonadal development of Scatophagus argus,we cloned the full-length cDNA sequence of antik-Müllerian hormone (AMH) from S.argus using the RACE technique,which is 2 324 bp in length (GenBank no.:KP718479).The open reading frame (ORF) of this gene spanned a region of 1 631 bp and encoded 543 amino acids.AMH of S.argus shared 71.16% similarity in sequence with that of Lateolabrax japonicus,while only 29.83% similarity with that of Danio rerio.S.alignment analysis revealed that S.argus AMH is closely related to Perciformes AMH,indicating AMH is conserved during evolution.The protein peptide contains a signal peptide (1-28 amino acids),an AMH-N domain (69-426 amino acids),and a TGF-β conserved domain (444-543 amino acids).Results of Real-time PCR showed that the mRNA level of AMH in testis of adult S.argus is significantly higher than that in other organs. AMH expression was also detected in liver and ovary.In addition,the mRNA expression of testis AMH at the development stages of I,II and III was kept at a high level,but was significantly decreased at the development stage of IV.According to the results of H.E staining,spermatogenesis happened in these stages,indicating an important role of AMH in testis development and the sperm production.In the ovary, AMH expression level in Ⅰ,Ⅱ oocyte development stages is low,while in the Ⅲ,Ⅳ stages,when the oocytes are rapidly growing and yolk are gradually accumulating,the AMH level is high.The results demonstrated that AMH is involved in the development and function maintaining of oocyte.Taken together, AMH played a significant role in the development and function of both testis and ovary.