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中华鲟中生殖细胞标记基因——死亡相关蛋白同源物的鉴定

Identification of a germ cell marker gene, the dead end homologue, in Chinese sturgeon Acipenser sinensis.

作者信息

Yang Xiaoge, Yue Huamei, Ye Huan, Li Chuangju, Wei Qiwei

机构信息

Institute of Hydrobiology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Wuhan 430072, China; Key Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture of China, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China.

Key Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture of China, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China; Freshwater Fisheries Research Center, Chinese Academy of Fisheries Science, Wuxi 214081, China.

出版信息

Gene. 2015 Mar 1;558(1):118-25. doi: 10.1016/j.gene.2014.12.059. Epub 2014 Dec 27.

Abstract

Dead end (dnd) encodes an RNA-binding protein that is essential for primordial germ cell (PGC) migration and gametogenesis in vertebrates. In this study, a Chinese sturgeon Acipenser sinensis dead end homologue, designated Asdnd, was identified and characterized. The full-length cDNA of Asdnd was 1630base pairs (bp) and encoded a peptide of 396 amino acid residues. Multiple sequence alignment showed that AsDnd shared six conserved regions of Dnd orthologs, including the RNA recognition motif. Phylogenetic analysis revealed that AsDnd was grouped with teleosts. By quantitative real-time PCR analysis, the Asdnd transcripts were found to originate from the maternal parent and be specifically expressed in gonads of immature Chinese sturgeons of both sexes. Fluorescent in situ hybridization indicated that Asdnd transcripts were restricted to germ cells. In the testis, Asdnd was abundant in spermatogonia and tended to gradually diminish in late spermatogenic stages, while strong signals were found in primary oocytes, as opposed to oogonia, in the ovary. Zebrafish PGCs were clearly visualized at 24h post-fertilization by co-injecting RFP-Asdnd 3' UTR and GFP-nos3 3' UTR mRNA, indicating that dnd 3' UTR has a conserved function among teleosts. Therefore, dnd could serve as a germ cell marker in Chinese sturgeon.

摘要

“死亡终端”(dnd)编码一种RNA结合蛋白,该蛋白对脊椎动物原始生殖细胞(PGC)的迁移和配子发生至关重要。在本研究中,鉴定并表征了中华鲟的一个“死亡终端”同源物,命名为Asdnd。Asdnd的全长cDNA为1630个碱基对(bp),编码一个由396个氨基酸残基组成的肽段。多序列比对显示,AsDnd与Dnd直系同源物共有六个保守区域,包括RNA识别基序。系统发育分析表明,AsDnd与硬骨鱼聚为一类。通过定量实时PCR分析发现,Asdnd转录本源自母本,且在未成熟的雌雄中华鲟性腺中特异性表达。荧光原位杂交表明,Asdnd转录本局限于生殖细胞。在睾丸中,Asdnd在精原细胞中丰富,在精子发生后期趋于逐渐减少,而在卵巢中,与卵原细胞相反,在初级卵母细胞中发现强信号。通过共注射RFP-Asdnd 3'UTR和GFP-nos3 3'UTR mRNA,在受精后24小时可清晰观察到斑马鱼的PGC,这表明dnd 3'UTR在硬骨鱼中具有保守功能。因此,dnd可作为中华鲟生殖细胞的标记物。

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