Development of a simplified and standardized protocol with potential for high-throughput for sperm cryopreservation in zebrafish Danio rerio
Development of a simplified and standardized protocol with potential for high-throughput for sperm cryopreservation in zebrafish Danio rerio
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DOI：
10.1016/j.theriogenology.2007.02.015
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发表时间：
2007-07-15
期刊：
影响因子：
2.8
通讯作者：
Tiersch, Terrence R.
中科院分区：
文献类型：
--
作者：
Yang, Huiping;Carmichael, Carrie;Tiersch, Terrence R.
精子冷冻保存为遗传资源的长期保存提供了可能。然而，目前的斑马鱼Danio rerio的方案是繁琐的，重现性差。我们的目标是通过简化从精子收集到解冻和使用的方法来促进冷冻保存的采用。首先，评价精子活化，当扩展剂的渗透压摩尔浓度>= 295-300 mOsmol/kg时，活动力完全被抑制。为了评价冷冻保护剂的毒性，将精子与5%、10%和15%浓度的二甲基亚砜（DMSO）、N，N-二甲基乙酰胺（DMA）、甲醇或甘油孵育。根据运动性，DMSO，DMA和甲醇（
Sperm cryopreservation offers potential for long-term storage of genetic resources. However, the current protocols for zebrafish Danio rerio are cumbersome and poorly reproducible. Our objective was to facilitate adoption of cryopreservation by streamlining methods from sperm collection through thawing and use. First, sperm activation was evaluated, and motility was completely inhibited when osmolality of the extender was >= 295-300 mOsmol/kg. To evaluate cryoprotectant toxicity, sperm were incubated with dimethyl sulfoxide (DMSO), N,N-dimethyl acetamide (DMA), methanol, or glycerol at 5, 10, and 15% concentrations. Based on motility, DMSO, DMA, and methanol (