title: Identification and sexually dimorphic expression of vasa isoforms in Dabry's sturgeon (Acipenser dabryanus), and functional analysis of vasa 3'-untranslated region.
abstract: Germ cells are set aside from somatic cells early in embryogenesis, and are responsible for transmitting genetic information through generations. Vasa is a highly conserved germ cell marker across animal phyla, and widely used to label primordial germ cells. Dabry's sturgeon is a rare and endangered species distributed solely in the Yangtze River basin. Here, seven vasa isoforms, named Advasa1-7, were isolated and characterized in Dabry's sturgeon. RT-PCR and western blot analyses revealed that vasa mRNA and protein were mainly restricted to the testis and ovary, but exhibited sexually dimorphic expression. Cellular and subcellular localization uncovered that Advasa mRNA and protein displayed mitotic and meiotic expression in females, and mainly showed mitotic expression in males; surprisingly, they exhibited both cytoplasmic and nuclear expression in the ovarian germ cells, while showing exclusively cytoplasmic expression in the testicular germ cells. By microinjecting chimeric RNA consisting of the red fluorescent protein coding region and the Advasa 3'-untranslated region into embryos of Dabry's sturgeon, zebrafish and medaka, we demonstrated that it had the ability to visualize primordial germ cells (PGCs) in Dabry's sturgeon and zebrafish but not in medaka. It seemed that the machinery of vasa 3'UTR RNA localization was conserved between Dabry's sturgeon and ostariophysan, while possibly changed during the divergence of euteleosts and ostariophysan. Finally, Dabry's sturgeon PGCs moved on the yolk ball, and migrated toward the genital ridge via mesenchyme. Taken together, these results provide new information for vasa expression pattern and function, and lay a foundation for PGC cryopreservation and conservation of Dabry's sturgeon.
authors.affiliation: Institute of Hydrobiology, Chinese Academy of Sciences, University of the 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.
authors.forename: Hua-Mei
authors.initials: HM
authors.lastname: Yue
authors.affiliation: Institute of Hydrobiology, Chinese Academy of Sciences, University of the 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: Xiao-Ge
authors.initials: XG
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. lcj@yfi.ac.cn.|Freshwater Fisheries Research Center, Chinese Academy of Fisheries Science, Wuxi, 214081, China. lcj@yfi.ac.cn.
authors.forename: Chuang-Ju
authors.initials: CJ
authors.lastname: Li
authors.affiliation: Institute of Hydrobiology, Chinese Academy of Sciences, University of the Chinese Academy of Sciences, Wuhan, 430072, China. weiqw@yfi.ac.cn.|Key Laboratory of Freshwater Biodiversity Conservation, Ministry of Agriculture of China, Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, 430223, China. weiqw@yfi.ac.cn.|Freshwater Fisheries Research Center, Chinese Academy of Fisheries Science, Wuxi, 214081, China. weiqw@yfi.ac.cn.
authors.forename: Qi-Wei
authors.initials: QW
authors.lastname: Wei
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002452:Cell Count; D002465:Cell Movement; D053487:DEAD-box RNA Helicases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D005260:Female; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005399:Fishes / Q000196:embryology / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000166:cytology / Q000378:metabolism; D006066:Gonads / Q000166:cytology / Q000378:metabolism; D008297:Male; D008845:Microinjections; D009990:Oryzias; D010802:Phylogeny; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D012727:Sex Characteristics; D015027:Zebrafish
chemical_list: D020413:3' Untranslated Regions; D029941:Fish Proteins; D018809:Proliferating Cell Nuclear Antigen; D020033:Protein Isoforms; D012333:RNA, Messenger; D053487:DEAD-box RNA Helicases
keywords: Dabry’s sturgeon; Dimorphic expression; Primordial germ cells; Vasa; Visualization