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当前位置: 首页 > 文章 > 中华鳖dazl基因克隆及在生殖细胞中的表达 水产学报 2019 (2) 400-409
Position: Home > Articles > The cloning and expression analysis of dazl in germ cells in the Chinese soft-shelled turtle(Pelodiscus sinensis) Journal of Fisheries of China 2019 (2) 400-409

中华鳖dazl基因克隆及在生殖细胞中的表达

作  者：
唐舟凯;张飘逸;储张杰;朱新平;李伟;吴栩灵;徐红艳
单  位：
浙江海洋大学水产学院;中国水产科学研究院珠江水产研究所农业农村部热带亚热带水产资源利用与养殖重点实验室
关键词：
中华鳖;dazl;生殖细胞;精子发生;卵子发生
摘  要：
为探索龟鳖类生殖细胞的发育分化机制,实验通过特异性引物克隆了中华鳖dazl基因的cDNA片段,长1 007 bp,其中包括5′端非编码区197 bp,3′端非编码区33 bp,开放阅读框777 bp,编码258个氨基酸。氨基酸序列比对显示其与西部锦龟Dazl同源性最高,达96%;与小鼠Dazl同源性达75%。荧光定量PCR分析结果显示,中华鳖dazl mRNA主要在精巢和卵巢中表达,在体细胞组织中仅检测到微量表达。冰冻切片原位杂交结果显示,中华鳖dazl mRNA在生殖细胞中特异表达,且在不同时期的生殖细胞中呈动态表达模式。在精巢中,中华鳖dazl mRNA在初级和次级精母细胞中表达最强,在精原干细胞中表达水平次之,在精子细胞中未检测到信号;在卵巢中,中华鳖dazl mRNA信号在初级卵母细胞胞质中均匀分布且在最早期的初级卵母细胞中信号最强,随着卵母细胞的增大,信号逐渐聚集并逐渐减弱。研究表明,dazl基因可能对中华鳖两性生殖细胞的发生具有重要的调控作用。
译  名：
The cloning and expression analysis of dazl in germ cells in the Chinese soft-shelled turtle(Pelodiscus sinensis)
作  者：
TANG Zhoukai;ZHANG Piaoyi;CHU Zhangjie;ZHU Xinping;LI Wei;WU Xuling;XU Hongyan;Fishery School of Zhejiang Ocean University;Key Laboratory of Tropical & Fishery Resource Application & Cultivation of Ministry of Agriculture and Rural Affaris,Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences;
单  位：
TANG Zhoukai%ZHANG Piaoyi%CHU Zhangjie%ZHU Xinping%LI Wei%WU Xuling%XU Hongyan%Fishery School of Zhejiang Ocean University%Key Laboratory of Tropical & Fishery Resource Application & Cultivation of Ministry of Agriculture and Rural Affaris,Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences
关键词：
Pelodiscus sinensis;;dazl;;germ cell;;spermatogenesis;;oogenesis
摘  要：
In this study, we used Pelodiscus sinensis as a model to study the germ cells differentiation in reptiles.Firstly, we cloned P. sinensis dazl cDNA fragment of 1 007 bp, containing a 197 bp 5′ untranslated region(UTR),a 33 bp 3′ UTR and an open reading frame(ORF) for 258 amino acid residues. The predicted P. sinensis Dazl was maximally 96% identical with Chrysemys picta bellii Dazl protein, 75% identical with Mus musculus Dazl protein.The RT-PCR result showed that the P. sinensis dazl mRNA was absent in all somatic tissues, but abundant in adult ovary and testis. Chemical in situ hybridization revealed that the P. sinensis dazl mRNA was exclusively expressed in germ cells but barely detected in somatic cells, and displayed a dynamic distribution expression pattern in germ cells during gametogenesis. In adult testis, the dazl mRNA signals were strongly dispersed in primary spermatocyte and secondary spermatocyte, weakly in spermatogonia, barely detected in spermatids. In adult ovary,the dazl mRNA signals were strong in early primary stages of oocytes and displayed a uniform distribution; as oocytes grew into larger size, the dazl mRNA signals were concentrated in the perinuclear cytoplasm and became weak as vitellogenesis further developed. In conclusion, the findings in this study suggested that dazl gene would play important roles in germ cells differentiation during gametogenesis in P. sinensis. The characterization of dazl gene in P. sinensis would provide the basis for further invstigations into reproductive development in turtles or reptiles.

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