为了解Sox14基因在拟穴青蟹胚胎发育和性腺发育过程中可能起到的调控作用,实验从青蟹性腺转录组数据库中得到全长为2558 bp的Sp-Sox14 cDNA序列,该序列编码一个包含427个氨基酸的蛋白,包含一个HMG-box.进化树分析表明,Sp-Sox14与其他节肢动物的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中的表达量最高.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他时期.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他发育阶段;而在精巢发育过程中,其表达量在成熟精子期(T3)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,在近孵化期定位于复眼附近,幼体孵出期则在头部仍有少量信号,暗示其与青蟹神经器官的形成以及体节附肢的发生有关.%In this study, a full-length 2558 bpSp-Sox14 cDNA sequence was obtained from Scylla paramamosain gonadal transcriptome database. The encoded protein consists of 427 amino acids and includes an HMG box do-main. Phylogenetic tree analysis showed that Sp-Sox14 was closely clustered with Sox14 of other arthropods. Quantitative real-time PCR displayed thatSp-Sox14 had different expression levels in different tissues from ma-ture male and female crabs with the highest expression in ovary and the heart of male crabs. During embryogenes-is,Sp-Sox14 expression was the highest in eye-pigment formation stage. During the different stages of ovarian de-velopment,Sp-Sox14 was expressed at the highest level in previtellogenesis stage (O2) and which was signific-antly higher than other stages. During the testis development, the expression level of Sp-Sox14 was higher in ma-ture sperm stage (T3) than in spermatocyte stage (T1) and spermatid stage (T2). It was speculated that Sp-Sox14 was involved in the early development of ovary and the process of sperm maturation. The whole mountin situ hy-bridization (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 was involved in the formation of the nervous organ as well as related to the occurrence of somite appendages.
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