title: Identification of a germ cell marker gene, the dead end homologue, in Chinese sturgeon Acipenser sinensis.
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.
authors.affiliation: 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.
authors.forename: Xiaoge
authors.initials: X
authors.lastname: Yang
authors.affiliation: 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.
authors.forename: Huamei
authors.initials: H
authors.lastname: Yue
authors.affiliation: 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.
authors.forename: Huan
authors.initials: H
authors.lastname: Ye
authors.affiliation: 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. Electronic address: lcj@yfi.ac.cn.
authors.forename: Chuangju
authors.initials: C
authors.lastname: Li
authors.affiliation: 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; Freshwater Fisheries Research Center, Chinese Academy of Fisheries Science, Wuxi 214081, China. Electronic address: weiqw@yfi.ac.cn.
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Wei
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015415:Biomarkers; D003001:Cloning, Molecular; D029941:Fish Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005854:Germ Cells / Q000378:metabolism; D017404:In Situ Hybridization, Fluorescence; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010473:Perciformes / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D016601:RNA-Binding Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D015027:Zebrafish
chemical_list: D015415:Biomarkers; D029941:Fish Proteins; D016601:RNA-Binding Proteins
keywords: Chinese sturgeon; Dead end; Fluorescent in situ hybridization; Primordial germ cell; qRT-PCR