中华鲟生殖细胞的标记基因研究新进展-长江水产研究所 长江水产研究所

中华鲟生殖细胞的标记基因研究新进展

  近期,课题组科研人员对中华鲟原始生殖细胞的迁移和配子发生至关重要的基因dnd进行了分离鉴定,并对其编码氨基酸结构及其在中华鲟生殖细胞中的表达进行了研究。研究论文在国际知名杂志《Gene》上发表。

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 Asdndwas 1630 base 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 transcriptswere restricted to germcells. 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 24 h 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 germcellmarker in Chinese sturgeon.

摘要翻译:Dead enddnd)编码一种RNA结合蛋白,对脊椎动物原始生殖细胞的迁移和配子发生至关重要。在本研究中,分离鉴定了中华鲟(Acipenser sinensisdead end的同源基因,即Asdnd。中华鲟dnd基因全长cDNA序列为1630 bp,编码396个氨基酸。通过氨基酸序列多重对比发现AsDnd与其他物种Dnd一样,具有包括RNA识别域在内的6个保守结构域。进化分析表明,中华鲟Dnd属于硬骨鱼类分支。qRT-PCR结果显示,Asdnd mRNA为母系遗传且仅在性腺组织中有表达。荧光原位杂交研究发现,Asdnd在生殖细胞中特异表达。在精巢中,Asdnd mRNA在精原细胞中表达最强,在后续的精子发生时期逐步减弱;在卵巢中,Asdnd mRNA在初级卵母细胞的表达较卵原细胞强。此外,通过显微共注射RFP-Asdnd 3′ UTR和GFP-nos3 3′ UTR mRNA,斑马鱼的原始生殖细胞(PGCs)能被成功标记,这表明dnd 3′ UTR在硬骨鱼中可能具有保守功能。因此,dnd 可以作为中华鲟生殖细胞的标记基因。