Abstract:The purpose of this study was to preliminarily investigate the role of Dmrta1(doublesex-and mab-3-related transcription factor A1) gene in gonad development in northern pike Esox lucius. In this study, Dmrta1 gene of northern pike cloned by RT-PCR had sequence length of 1408 bp, encoding 433 amino acids, the predicted molecular weight of 36.047 ku, and the theoretical isoelectric point (PI) of 5.94. The gene structure and the evolutionary relationship of species were carried out using the SMART, Swiss-Model and MEGA software to be evaluated.Protein structural prediction based on the putative amino acid sequence of the Dmrta1 revealed that Dmrta1 gene of E. lucius contained a DM domain and a DMA domain, indicating that it was more phylogenetically related to Oncorhynchus mykiss and O. nerka. The homology of amino acid sequence of Dmrta1 gene in E. lucius was above 75% with O. kisutch,O. nerka,O. mykiss and Salmo salar, respectively. Real-time PCR analysis showed that Dmrta1 gene was expressed in male and female gonads, gills, meat and other tissues. The expression of Dmrta1 gene was significantly higher in testis than that in ovary (P<0.05).At the age of 180 days and 320 days, this gene was highly expressed in the gonads and gills of male E. lucius .The findings showed that the expression level of Dmrta1 gene in the testis was increased during development, indicating that it plays an important role in the maturation of testis. The Dmrta1 gene may play an important role in the formation of gills and the promotion of male sex differentiation and function maintenance.
[1]Burtis K C, Baker B S. Drosophila double sex gene controls somatic sexual differentiation by producing alternatively spliced mRNAs encoding related sex-specific polypeptides[J].Cell,1989,56(6):997-1010. [2]Guan G J, Kobayashi T, Nagahama Y. Sexually dimorphic expression of two types of DM (doublesex/Mab-3)-domain genes in a teleost fish,the tilapia (Oreochromis niloticus)[J].Biochemical and Biophysical Research Communications,2000,272(3):662-666. [3]Raymond C S, Kettlewell J R, Hirsch B, et al. Expression of Dmrt1 in the genital ridge of mouse and chicken embryos suggests a role in vertebrate sexual development[J].Developmental Biology,1999,215(2):208-220. [4]Raymond C S, Shamu C E, Shen M M, et al. Evidence for evolutionary conservation of sex-determining genes[J].Nature,1998,391(6668):691-695. [5]Smith C A, McClive P J, Western P S, et al. Conservation of a sex-determining gene[J].Nature,1999,402(6762):601-602. [6]任莉莉,程汉华,郭一清,等.两栖类、爬行类和鸟类存在一个新的Dmrt基因家族[J].科学通报,2001,46(14):1187-1190. [7]Volff J N, Zarkower D, Bardwell V J, et al. Evolutionary dynamics of the DM domain gene family in metazoans[J].Journal of Molecular Evolution,2003,57:S241-S249. [8]Balciuniene J, Bardwell V J, Zarkower D. Mice mutant in the DM domain gene Dmrt4 are viable and fertile but have polyovular follicles[J].Molecular and Cellular Biology,2006,26(23):8984-8991. [9]董晓丽,陈松林.半滑舌鳎Dmrt4基因DNA全序列的克隆与表达分析[J].中国水产科学,2013,20(3):499-505. [10]江东能,彭友幸,奥马尔·法鲁克·穆斯塔法,等.金钱鱼Dmrt4基因的克隆及表达分析[J].广东海洋大学学报,2019,39(1):7-13. [11]Cejko B I, Krejszeff S, Z·arski D, et al. Effect of carp pituitary homogenate (CPH) and sGnRHa (Ovaprim) on northern pike (Esox lucius) spermiation stimulation and its effect on quantity and quality of sperm[J].Animal Reproduction Science,2018,193:217-225. [12]任慕莲,郭焱,张人铭,等.中国额尔齐斯河鱼类资源及渔业[M].乌鲁木齐:新疆科技卫生出版社,2002:86-87 [13]齐遵利,张秀文,韩叙.温度对白斑狗鱼早期发育和生长的影响[J].水产科学,2010,29(12):708-710. [14]Sunde J, Larsson P, Forsman A. Adaptations of early development to local spawning temperature in anadromous populations of pike (Esox lucius)[J].BMC Evolutionary Biology,2019,19:148. [15]李飞,胡琼,张俊杰.17α-甲基睾丸酮诱导白斑狗鱼性逆转初步研究[J].淡水渔业,2018,48(1):3-8. [16]刘良国,赵俊,崔淼.鱼类的性别决定与人工控制[J].水产科学,2003,22(2):42-45. [17]季代丽,李洁,刘先英,等.黑斑蛙Dmrt1基因的克隆及在不同组织中的表达[J].应用与环境生物学报,2005,11(2):198-201. [18]周丽青,杨爱国,王清印,等.虾夷扇贝不同性别类型2个Dmrt基因DM结构域分析[J].海洋科学,2015,39(3):19-25. [19]吴启迪,谭月,王靖天,等.雌二醇刺激对菲律宾蛤仔Dmrt基因表达的影响[J].大连海洋大学学报,2019,34(3):362-369. [20]Yamaguchi A, Lee K H, Fujimoto H, et al. Expression of the DMRT gene and its roles in early gonadal development of the Japanese pufferfish Takifugu rubripes[J].Comparative Biochemistry and Physiology Part D:Genomics and Proteomics,2006,1(1):59-68. [21]Winkler C, Hornung U, Kondo M, et al. Developmentally regulated and non-sex-specific expression of autosomal dmrt genes in embryos of the Medaka fish (Oryzias latipes)[J].Mechanisms of Development,2004,121(7/8):997-1005. [22]Wen A Y, You F, Tan X G, et al. Expression pattern of dmrt4 from olive flounder (Paralichthys olivaceus) in adult gonads and during embryogenesis[J].Fish Physiology and Biochemistry,2009,35(3):421-433.