DNA methylation stability in fish spermatozoa upon external constraint: Impact of fish hormonal stimulation and sperm cryopreservation
DNA methylation stability in fish spermatozoa upon external constraint: Impact of fish hormonal stimulation and sperm cryopreservation
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DOI：
10.1002/mrd.23297
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发表时间：
2020-01-01
影响因子：
2.5
通讯作者：
Labbe, Catherine
中科院分区：
文献类型：
--
作者：
Depince, Alexandra;Gabory, Anne;Labbe, Catherine
高度分化的成熟精子不仅携带遗传信息，还携带将传递给胚胎的表观遗传信息。DNA甲基化是一种与精子核致密化、基因沉默和胚胎基因表达预模式化相关的表观遗传因子。因此，该标记对生殖生物技术的稳定性是动物生产中的一个主要问题。本工作探讨了激素诱导的精子和精子冷冻保存的影响，在两个鲤科鱼类，金鱼（鲫鱼）和斑马鱼（斑马鱼），使用LUMA的甲基化测定。我们发现，虽然金鱼激素治疗确实增加了精子产量，但它并没有改变精子的整体DNA甲基化。从同一男性身上重复收集的不同精子样本在2个月内也显示出相同的整体DNA甲基化水平。同样，全球DNA甲基化没有受到影响后，冷冻保存金鱼精子与甲醇，而效率较低的冷冻保护剂（二甲基亚砜和1，2-丙二醇）减少DNA甲基化。与此相反，冷冻保存的斑马鱼精子与甲醇诱导轻微，但显着的，增加全球DNA甲基化。在不太紧凑的细胞核，即金鱼鳍体细胞，冷冻保存没有改变全球DNA甲基化，无论选择的冷冻保护剂。总之，整体DNA甲基化是一个强大的参数，如激素诱导精子和精子冷冻保存生物技术，但它可以改变时，最佳的精子操作条件不满足。
Highly differentiated mature spermatozoa carry not only genetic but also epigenetic information that is to be transmitted to the embryo. DNA methylation is one epigenetic actor associated with sperm nucleus compaction, gene silencing, and prepatterning of embryonic gene expression. Therefore, the stability of this mark toward reproductive biotechnologies is a major issue in animal production. The present work explored the impact of hormonal induction of spermiation and sperm cryopreservation in two cyprinids, the goldfish (Carassius auratus) and the zebrafish (Danio rerio), using LUminometric Methylation Assay (LUMA). We showed that while goldfish hormonal treatment did increase sperm production, it did not alter global DNA methylation of spermatozoa. Different sperm samples repeatedly collected from the same males for 2 months also showed the same global DNA methylation level. Similarly, global DNA methylation was not affected after cryopreservation of goldfish spermatozoa with methanol, whereas less efficient cryoprotectants (dimethylsulfoxide and 1,2-propanediol) decreased DNA methylation. In contrast, cryopreservation of zebrafish spermatozoa with methanol induced a slight, but significant, increase in global DNA methylation. In the less compact nuclei, that is, goldfish fin somatic cells, cryopreservation did not change global DNA methylation regardless of the choice of cryoprotectant. To conclude, global DNA methylation is a robust parameter with respect to biotechnologies such as hormonal induction of spermiation and sperm cryopreservation, but it can be altered when the best sperm manipulation conditions are not met.