title: Sexual dimorphic expression of dnd in germ cells during sex reversal and its requirement for primordial germ cell survival in protogynous hermaphroditic grouper.
abstract: Dead end (dnd), vertebrate-specific germ cell marker, had been demonstrated to be essential for primordial germ cell (PGC) migration and survival, and the link between PGC number and sex change had been revealed in some teleost species, but little is known about dnd in hermaphroditic vertebrates. In the present study, a protogynous hermaphroditic orange-spotted grouper (Epinephelus coioides) dnd homologue (Ecdnd) was identified and characterized. Quantitative real-time PCR and in situ hybridization analysis revealed a dynamic and sexually dimorphic expression pattern in PGCs and germ cells of gonads. During sex changing, the Ecdnd transcript sharply increased in early transitional gonad, reached the highest level at late transitional gonad stage, and decreased after testis maturation. Visualization of zebrafish PGCs by injecting with RFP-Ecdnd-3'UTR RNA and GFP-zfnanos3-3'UTR RNA confirmed importance of Ecdnd 3'UTR for the PGC distribution. In addition, knockdown of EcDnd by using antisense morpholinos (MO) caused the ablation of PGCs in orange-spotted grouper. Therefore, the current data indicate that Ecdnd is essential for PGCs survival and may serve as a useful germ cell marker during gametogenesis in hermaphroditic grouper.
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate University of the Chinese Academy of Sciences, Wuhan 430072, China.
authors.forename: Zhi-Hui
authors.initials: ZH
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate University of the Chinese Academy of Sciences, Wuhan 430072, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate University of the Chinese Academy of Sciences, Wuhan 430072, China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Li
authors.affiliation: State Key Laboratory of Biocontrol, Guangdong Province Key Laboratory for Aquatic Economic Animals, Institute of Aquatic Economic Animals, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, China.
authors.forename: Xiao-Chun
authors.initials: XC
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Biocontrol, Guangdong Province Key Laboratory for Aquatic Economic Animals, Institute of Aquatic Economic Animals, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, China.
authors.forename: Shui-Sheng
authors.initials: SS
authors.lastname: Li
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate University of the Chinese Academy of Sciences, Wuhan 430072, China. Electronic address: wangyang@ihb.ac.cn.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate University of the Chinese Academy of Sciences, Wuhan 430072, China. Electronic address: jfgui@ihb.ac.cn.
authors.forename: Jian-Fang
authors.initials: JF
authors.lastname: Gui
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D005260:Female; D029941:Fish Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000166:cytology* / Q000378:metabolism; D006066:Gonads / Q000196:embryology / Q000378:metabolism*; D057850:Hermaphroditic Organisms / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D017403:In Situ Hybridization; D008297:Male; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D012727:Sex Characteristics; D012733:Sex Differentiation; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*
chemical_list: D029941:Fish Proteins
keywords: Epinephelus coioides; Morpholino; PGC; Sex change; dnd