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泥蟹 Scylla paramamosain 性腺差异表达 SoxB2 基因的特征分析。

Characterization of gonad differentially expressed SoxB2 genes in mud crab Scylla paramamosain.

机构信息

Key Laboratory of Healthy Mariculture for the East China Sea, Ministry of Agriculture and Rural Affairs, Fisheries College, Jimei University, Xiamen 361021, China.

Key Laboratory of Marine Biotechnology of Fujian Province, Institute of Oceanology, College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China.

出版信息

Gene. 2020 May 25;740:144507. doi: 10.1016/j.gene.2020.144507. Epub 2020 Feb 26.

Abstract

Members of sox gene family play critical roles in development, and some of them have crucial functions in sexual dimorphism. To understand the role of two SoxB2 genes, Sox14b and Sox21 of mud crab Scylla paramamosain, the full-length 1939 bp SpSox14b cDNA sequence and 861 bp SpSox21 cDNA sequence were obtained from the crab's transcriptome database, which encode 397 and 259 amino acids respectively. The results of sq-PCR showed that SpSox14b was expressed in all tissues, while SpSox21 was only expressed in the testis and brain. qRT-PCR showed that the expression level of SpSox14b in ovary was significantly higher than that of testis, and during the gonad development its expression was the highest in O2 (previtellogenesis) stage. The expression level of SpSox21 in testis was much higher than in brain, and was significantly higher in T3 (the mature sperm stage) than in other stages of testis development. Meanwhile, in different stages of larval development, SpSox21 was low expressed in zoea, then increased significantly in megalopa. Therefore we speculated that SpSox14b and SpSox21 may play different roles in the gonad development of mud crab, especially SpSox21 may be involved in the development and maintenance of testis. The expression level of SpSox14b and SpSox21 during the eye-pigment formation was significantly higher than that in other embryonic development stages, the results of whole-mount in situ hybridization showed that SpSox14b and SpSox21 were mainly located near the head and the compound eyes in eye-pigment formation stage and hatching. It suggested that they may be involved in the formation of brain nerves and are related to the regulation of body segments, and play different roles in sexual dimorphism.

摘要

Sox 基因家族的成员在发育过程中发挥着关键作用,其中一些在性别二态性中具有至关重要的功能。为了了解泥蟹 Scylla paramamosain 中两个 SoxB2 基因 Sox14b 和 Sox21 的作用,从蟹的转录组数据库中获得了全长 1939bp 的 SpSox14b cDNA 序列和 861bp 的 SpSox21 cDNA 序列,分别编码 397 和 259 个氨基酸。sq-PCR 结果表明,SpSox14b 在所有组织中均有表达,而 SpSox21 仅在睾丸和脑中表达。qRT-PCR 结果显示,SpSox14b 在卵巢中的表达水平明显高于睾丸,在卵巢发育过程中,O2(卵黄发生前)期表达水平最高。SpSox21 在睾丸中的表达水平明显高于脑中,在睾丸发育的 T3(成熟精子期)期明显高于其他发育阶段。同时,在不同的幼虫发育阶段,SpSox21 在幼体中低表达,在大眼幼虫中显著增加。因此,我们推测 SpSox14b 和 SpSox21 可能在泥蟹性腺发育中发挥不同的作用,特别是 SpSox21 可能参与睾丸的发育和维持。SpSox14b 和 SpSox21 在眼色素形成过程中的表达水平明显高于其他胚胎发育阶段,整体原位杂交结果表明,SpSox14b 和 SpSox21 主要位于头部和复眼附近在眼色素形成和孵化阶段。这表明它们可能参与脑神经的形成,并与身体节段的调节有关,在性别二态性中发挥不同的作用。

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