Solubilization of Mouse Sperm Chromatin for Sequencing Analyses Using a Chaperon Protein
Solubilization of Mouse Sperm Chromatin for Sequencing Analyses Using a Chaperon Protein
复制标题
使用伴侣蛋白溶解小鼠精子染色质进行测序分析
DOI：
10.1007/978-1-0716-2724-2_11
复制
发表时间：
2022
影响因子：
--
通讯作者：
Yuki Okada
中科院分区：
文献类型：
--
作者：
Yuko Fukuda;Keishi Shintomi;Kosuke Yamaguchi;Yasuhiro Fujiwara;Yuki Okada
精子染色质致密化是精子获得生育力的生理基础。然而，这种由鱼精蛋白组成的独特结构使我们无法溶解染色质，因为它对超声和通常用于体细胞中染色质片段化的酶具有抗性。即使当强烈的酶处理，它似乎只是溶解精子染色质的某些部分，大概是因为异质性。为了克服这个问题，我们以前开发了一种方法来处理精子与重组核质蛋白，在受精胚胎中的鱼精蛋白去除剂，然后超声处理。核质蛋白处理显着增加精子染色质溶解的效率，而需要相对大量的重组核质蛋白。本文介绍了一种改进的核质蛋白法，该方法重组蛋白量少，反应时间短。
Sperm chromatin compaction is physiologically essential for sperm to acquire the fertility. However, this unique structure composed of protamines makes us unable to solubilize the chromatin due to its resistance to sonication and enzymes usually used for chromatin fragmentation in somatic cells. Even when intense enzymatic treatment is applied, it appears to solubilize only certain portions of sperm chromatin presumably because of the heterogeneous properties. To overcome this issue, we previously developed a method to treat the sperm with recombinant nucleoplasmin, a protamine remover in fertilized embryos, followed by sonication. The nucleoplasmin treatment dramatically increased the efficiency of sperm chromatin solubilization, while a relatively large amount of recombinant nucleoplasmin was required. Here, we describe an improvement of nucleoplasmin method with a less amount of recombinant protein and a shorter reaction time.