从小鼠及其他啮齿动物中分离精子细胞核和核基质。
Isolation of sperm nuclei and nuclear matrices from the mouse, and other rodents.
作者信息
Ward W Steven
机构信息
Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI, USA.
出版信息
Methods Mol Biol. 2013;927:437-44. doi: 10.1007/978-1-62703-038-0_37.
Abstract
The isolation of mammalian sperm heads from their tails is complicated by the relatively high density of the tails, but facilitated by the fact that protamine condensation of the sperm chromatin and the insolubility of the perinuclear theca make the sperm nucleus stable in sodium dodecyl sulfate. Two methods are described for the isolation of rodent sperm nuclei using sucrose step gradients in which the sperm nuclei are only centrifuged one time, minimizing potential damage by mechanical stress.
摘要
从尾部分离哺乳动物的精子头部较为复杂，因为尾部的密度相对较高，但精子染色质的鱼精蛋白凝聚以及核周膜的不溶性使精子核在十二烷基硫酸钠中保持稳定，这有助于分离。本文描述了两种使用蔗糖阶梯梯度分离啮齿动物精子核的方法，其中精子核仅离心一次，将机械应力造成的潜在损伤降至最低。
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本文引用的文献
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Sperm-derived histones contribute to zygotic chromatin in humans.精子来源的组蛋白对人类合子染色质有贡献。
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Mouse spermatozoa contain a nuclease that is activated by pretreatment with EGTA and subsequent calcium incubation.小鼠精子含有一种核酸酶，该酶可通过用乙二醇双（2-氨基乙基醚）四乙酸（EGTA）预处理并随后进行钙孵育来激活。
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