缢蛏2个Dmrt5基因鉴定及其在性腺发育中的表达分析

IDENTIFICATION OF TWO DMRT5 GENES AND THEIR EXPRESSION ANALYSIS IN GONADAL DEVELOPMENT AND DIFFERENT STRESSES OF SINONOVACULA CONSTRICTA

中文关键词: 缢蛏  ScDmrt5-1  ScDmrt5-2  性腺发育  表达

英文关键词: Sinonovacula constricta  Gonadal development  ScDmrt5-1  ScDmrt5-2  Expression

中文摘要:

      Dmrt5是Dmrt基因家族的重要一员,是与动物性别决定、性腺发育、器官功能维持等密切相关的转录因子。为探讨缢蛏(Sinonovacula constricta) Dmrt5基因(ScDmrt5)在性腺发育中的作用,本研究克隆、鉴定了2个ScDmrt5基因,并研究了其在性腺不同发育时期的表达特征。ScDmrt5-1和ScDmrt5-2分别编码389、374个氨基酸,均具备DM和DMA保守结构域；在性成熟期,ScDmrt5-1和ScDmrt5-2在鳃、外套膜等组织中均有表达,并在精巢中的表达量显著高于卵巢(P＜0.05)；在性腺不同发育时期,ScDmrt5-1基因的表达量随着精巢不断发育成熟逐步升高,在成熟期表达量最高(P＜0.05),而在卵巢中的表达趋势与之相反；ScDmrt5-2基因表达量也随着精巢的不断发育成熟逐步升高,而在卵巢各个发育时期的表达量均较低且无显著差异。ScDmrt5-1和ScDmrt5-2蛋白均主要定位在精巢的精原细胞、精母细胞、精细胞和精子及卵巢的卵原细胞、卵母细胞和成熟卵细胞中。综上表明,ScDmrt5-1和ScDmrt5-2可能主要参与缢蛏精巢的发育及功能维持,并可能在呼吸、神经感知等发挥重要作用。

英文摘要:

      Dmrt5, an important member of the Dmrt gene family, is a transcription factor closely related to sex determination, gonad development and organ function maintenance in animals. In order to investigate the roles of Dmrt5 genes (ScDmrt5) in gonadal development of Sinonovacula constricta, the full-length cDNA sequences of the two ScDmrt5 were cloned and identified, and the expression patterns of ScDmrt5 genes/proteins during gonadal development were investigated. Based on sequence comparison and phylogenetic analysis, the two genes were named ScDmrt5-1 and ScDmrt5-2, encoding 389 and 374 amino acids, respectively, both of which possessed DM and DMA conserved structural domains. In the gonadal mature stage, ScDmrt5-1 and ScDmrt5-2 were all expressed in seven tissues, and the expression level in the testis was significantly higher than that in the ovary (P＜0.05). During gonadal development, the expression level of ScDmrt5-1 gradually increased with the development of the testis, and the expression level was highest in the mature stage (P＜0.05), while the expression trend was opposite in the ovary. The expression level of ScDmrt5-2 gradually increased with the development of testis, but the expression level was all lower and without significant difference among different developmental stages of ovary. The immunofluorescence results showed that ScDmrt5-1 and ScDmrt5-2 proteins were mainly located in the germ cells of male and female gonads during gonadal development, such as spermatogonia, spermatocytes and mature sperm of testis, and oocytes, oocytes and mature oocytes of ovary. In conclusion, ScDmrt5-1 and ScDmrt5-2 may be mainly involved in the development and functional maintenance of testis, and may be also involved in respiration, neuroperception and so on.