文章摘要
拟穴青蟹Sp-Sox14基因在胚胎发育和性腺发育过程中的表达研究
Expression analysis of Sp-Sox14 gene during embryonic and gonadal development in Scylla paramamosain
投稿时间:2017-02-12  修订日期:2017-04-09
DOI:
中文关键词: 拟穴青蟹  Sp-Sox14  胚胎发育  性腺发育  基因表达
英文关键词: Scylla paramamosain  Sp-Sox14  embryogenesis  gonadal development  gene expression
基金项目:国家自然科学基金
作者单位E-mail
梁可蓥 集美大学水产学院 1832353063@qq.com 
张子平 福建农林大学动物科学学院  
邹志华 集美大学水产学院  
王艺磊 集美大学水产学院 ylwang@jmu.edu.cn 
廖佳倩 福建农林大学动物科学学院  
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中文摘要:
      Sox基因家族是由众多编码具有HMG-box(high mobility group box)保守基序的基因构成的超基因家族,在性别决定与分化、早期胚胎发育、神经系统的发育和分化、软骨形成、血细胞形成和晶状体形成等多个过程起着关键作用。Sox14基因属于Sox基因家族中的B2亚族,作为转录阻遏物,与SoxB1亚族拮抗从而促进神经细胞分化。本研究从拟穴青蟹(Scylla paramamosain)性腺转录组数据库中得到全长为2558bp的Sp-Sox14 cDNA序列,该序列编码一个427个氨基酸的蛋白,包含一个HMG-box。进化树分析表明,Sp-Sox14与其它节肢动物的Sox14亲缘关系较近。实时定量PCR结果显示,Sp-Sox14在成蟹雌雄各组织中均有不同程度的表达,其中在卵巢和雄蟹心脏的表达量最高。在胚胎发育过程中,Sp-Sox14在复眼色素形成期的表达量显著高于其它时期。在性腺不同发育阶段,Sp-Sox14在卵黄发生前期(O2)的表达量显著高于卵巢其它发育阶段;而在精巢发育过程中,其表达量在成熟精子期(T3)显著高于精母细胞期(T1)和精子细胞期(T2)。推测其参与卵巢的前期发育以及精子成熟等过程。整胚原位杂交结果显示,Sp-Sox14在青蟹胚胎复眼色素形成期阳性信号定位于头部以及鄂足附近,在近孵化期位于定位于复眼附近,幼体孵出期则在头部以及附肢附近仍有少量信号,暗示其与青蟹神经系统的形成以及体节附肢的发生有关。
英文摘要:
      Members of the SOX family of transcription factors are characterized by the presence of a DNA-binding HMG (high mobility group) domain. The Sox proteins play critical roles in several processes, including sex determination, embryogenesis, central nervous system development, chondrogenesis, and the formation of blood cells and lens. The Sp-Sox14 gene belongs to group of B2 Sox genes and acts as a transcriptional repressor, which antagonizes SoxB1 and promotes neuronal differentiation. In this study, a full-length 2558bp Sp-Sox14 cDNA sequence was obtained from Scylla paramamosain gonadal transcriptome database. The encoded protein consists of 427 amino acids and includes an HMG box domain. Phylogenetic tree analysis showed that Sp-Sox14 was closely clustered with Sox14 of other arthropods. Quantitative real-time PCR displayed that Sp-Sox14 have different expression levels in different tissues from mature male and female crabs with the highest expression in ovary and the heart of male crabs. During embryogenesis, Sp-Sox14 expression was highest in eye-pigment formation stage. During the different stages of ovarian development, Sp-Sox14 was expressed at the highest level in previtellogenesis stage (O2) and was significantly higher than other stages. During the testis development, the expression level of Sp-Sox14 was higher in mature sperm stage (T3) than in spermatocyte stage (T1) and spermatid stage (T2). It was speculated that Sp-Sox14 is involved in the early development of ovary and the process of sperm maturation. The whole mount in situ hybridization (WISH) results indicated that the positive signals of Sp-Sox14 were mainly distributed in the head and near the maxillipede at eye-pigment formation stage and near the compound eyes at prehatching stage. A small number of Sp-Sox14 positive signals were still present in the head and near the appendages at hatching stage. The results of WISH implied that Sp-Sox14 is involved in the formation of the nervous system as well as related to the occurrence of somite appendages.
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