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精子DNA损伤和卵母细胞修复能力对鳟鱼发育的影响。

Impact of sperm DNA damage and oocyte-repairing capacity on trout development.

作者信息

Fernández-Díez C, González-Rojo S, Lombó M, Herráez M P

机构信息

Department of Molecular BiologyFaculty of Biology, University of León, León, Spain.

Department of Molecular BiologyFaculty of Biology, University of León, León, Spain

出版信息

Reproduction. 2016 Jul;152(1):57-67. doi: 10.1530/REP-16-0077. Epub 2016 Apr 12.

Abstract

Zygotic repair of paternal DNA is essential during embryo development. In spite of the interest devoted to sperm DNA damage, its combined effect with defect-repairing oocytes has not been analyzed. Modification of the breeding season is a common practice in aquaculture. This practice reduces developmental success and could affect the both factors: sperm DNA integrity and oocyte repair capacity. To evaluate the maternal role, we analyzed the progeny outcome after fertilizing in-season trout oocytes with untreated and with UV-irradiated sperm. We also analyzed the offspring obtained out of season with untreated sperm. The analysis of the number of lesions in 4 sperm nuclear genes revealed an increase of 1.22-11.18 lesions/10 kb in out-of-season sperm, similar to that obtained after sperm UV irradiation (400 µW/cm(2)5 min). Gene expression showed in out-of-season oocytes the overexpression of repair genes (ogg1, ung, lig3, rad1) and downregulation of tp53, indicating an enhanced repairing activity and reduced capacity to arrest development upon damage. The analysis of the progeny in out-of-season embryos revealed a similar profile tolerant to DNA damage, leading to a much lower apoptotic activity at organogenesis, lower hatching rates and increased rate of malformations. The effects were milder in descendants from in-season-irradiated sperm, showing an enhanced repairing activity at epibolia. Results point out the importance of the repairing machinery provided by the oocyte and show how susceptible it is to environmental changes. Transcripts related to DNA damage signalization and repair could be used as markers of oocyte quality.

摘要

在胚胎发育过程中,父本DNA的合子修复至关重要。尽管人们对精子DNA损伤颇感兴趣,但尚未分析其与缺陷修复卵母细胞的联合效应。改变繁殖季节是水产养殖中的常见做法。这种做法会降低发育成功率,并可能影响两个因素:精子DNA完整性和卵母细胞修复能力。为了评估母体的作用,我们分析了用未处理的和经紫外线照射的精子使季节性鲑鱼卵母细胞受精后的子代结果。我们还分析了用未处理的精子在非繁殖季节获得的后代。对4个精子核基因中的损伤数量进行分析发现,非繁殖季节精子中的损伤数量增加了1.22 - 11.18个损伤/10 kb,与精子紫外线照射后(400 μW/cm² 5分钟)的情况相似。基因表达显示,在非繁殖季节的卵母细胞中,修复基因(ogg1、ung、lig3、rad1)过表达,tp53下调,表明修复活性增强,损伤时阻止发育的能力降低。对非繁殖季节胚胎中的子代分析显示,其对DNA损伤具有相似的耐受性,导致器官发生时的凋亡活性低得多、孵化率低以及畸形率增加。在繁殖季节受照射精子的后代中,这些影响较为轻微,在囊胚期显示出增强的修复活性。结果指出了卵母细胞提供的修复机制的重要性,并表明其对环境变化的敏感性。与DNA损伤信号传导和修复相关的转录本可作为卵母细胞质量的标志物。

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