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鉴定黄颡鱼 H-P-G 轴中的生殖相关基因和途径,并分析畸形和正常雌核发育卵巢中这些基因表达的差异。

Identification of reproduction-related genes and pathways in the Culter alburnus H-P-G axis and characterization of their expression differences in malformed and normal gynogenetic ovaries.

机构信息

Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture, Key Laboratory of Freshwater Aquaculture Genetic and Breeding of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou, Zhejiang, China.

出版信息

Fish Physiol Biochem. 2021 Feb;47(1):1-20. doi: 10.1007/s10695-020-00859-9. Epub 2020 Nov 6.

Abstract

This study applied RNA-seq technology to discover reproduction-related genes and pathways in female topmouth culter brain (including pituitary) and ovarian tissues. In functional analysis, 2479 and 2605 unigenes in the brain and ovary tissue were assigned to the "reproductive process" subcategory in addition to the 2660 and 2845 unigenes assigned to the "reproduction" subcategory. Twenty-three complete cDNA sequences were identified through the different gene expression (DGE) approach from five reproduction-related pathways (MAPK signaling pathway, neuroactive ligand-receptor interaction pathway, gonadotropin-releasing hormone signaling pathway, oocyte meiosis pathway, and steroid biosynthesis pathway). The expression levels of 16 candidate genes using qPCR in this study were in accordance with the results of transcriptome analysis. In addition, the expression levels of the FSH, 3β-HSD, PGR, and NPYR genes in malformed gynogenetic ovaries were considerably low, which was consistent with the progress of oocytogenesis in the ovaries of topmouth culter. The high expression of these four genes in the ovaries of normal topmouth culter suggested they might involve in the preparation for the shift of oogenesis to ovulation. Hence, our work identified a set of annotated gene products that are candidate factors affecting reproduction in the topmouth culter H-P-G axis. These results could be essential for further research in functional genomics and genetic editing for topmouth culter reproduction.

摘要

本研究应用 RNA-seq 技术在雌性大口黑鲈脑(包括脑垂体)和卵巢组织中发现与生殖相关的基因和通路。在功能分析中,脑和卵巢组织中分别有 2479 个和 2605 个基因被分配到“生殖过程”亚类,而分别有 2660 个和 2845 个基因被分配到“生殖”亚类。通过五个与生殖相关的通路(MAPK 信号通路、神经活性配体-受体相互作用通路、促性腺激素释放激素信号通路、卵母细胞减数分裂通路和类固醇生物合成通路)的不同基因表达(DGE)方法,鉴定出 23 个完整的 cDNA 序列。本研究中使用 qPCR 检测的 16 个候选基因的表达水平与转录组分析结果一致。此外,在畸形雌核发育卵巢中,FSH、3β-HSD、PGR 和 NPYR 基因的表达水平明显较低,这与大口黑鲈卵巢中卵母细胞发生的进程一致。这四个基因在正常大口黑鲈卵巢中的高表达表明,它们可能参与了卵母细胞从发生到排卵的转变的准备过程。因此,我们的工作鉴定了一组注释基因产物,这些产物可能是影响大口黑鲈 H-P-G 轴生殖的候选因素。这些结果对于大口黑鲈生殖的功能基因组学和遗传编辑的进一步研究可能是至关重要的。

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