Analysis of Meiotic Sister Chromatid Cohesion in Caenorhabditis elegans.
Analysis of Meiotic Sister Chromatid Cohesion in Caenorhabditis elegans.
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秀丽隐杆线虫减数分裂姐妹染色单体凝聚力分析。
DOI：
10.1007/978-1-4939-6545-8_5
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Severson,AaronF
中科院分区：
文献类型：
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作者：
Severson,AaronF
在有性生殖生物中，健康配子（精子和卵子）的形成需要减数分裂姐妹染色单体凝聚（SCC）的正确建立和释放。SCC系链从减数分裂前S期的形成开始复制姐妹篇，直到减数分裂I和II后期逐渐去除凝聚力，允许同源物分离，然后是姐妹篇。减数分裂凝聚力的建立或释放的缺陷导致染色体分离错误，从而导致非整倍体配子和不存活胚胎的形成。线虫秀丽隐杆线虫由于其遗传上的易处理性和雌雄同体性腺的优良细胞学特性，是研究减数分裂姐妹染色单体凝聚的一个有吸引力的模型。此外，在减数分裂SCC的建立或维持缺陷的突变体，但产生丰富的配子，允许分析的染色体分离的模式。在这里，我描述了两种方法来分析减数分裂凝聚在C。第一种方法依赖于细胞学来检测和量化SCC中的缺陷。第二种方法依赖于PCR和限制性内切酶来鉴定由于异常减数分裂染色体分离而遗传了不正确的染色体互补的胚胎。这两种方法都足够敏感，可以识别罕见的错误，并且足够精确，可以揭示以不同方式扰乱减数分裂SCC的突变所产生的独特表型。这些测定的稳健性和定量性应该加强不同减数分裂突变体的表型比较，并增强不同研究者生成的数据的可重复性。
In sexually reproducing organisms, the formation of healthy gametes (sperm and eggs) requires the proper establishment and release of meiotic sister chromatid cohesion (SCC). SCC tethers replicated sisters from their formation in premeiotic S phase until the stepwise removal of cohesion in anaphase of meiosis I and II allows the separation of homologs and then sisters. Defects in the establishment or release of meiotic cohesion cause chromosome segregation errors that lead to the formation of aneuploid gametes and inviable embryos. The nematodeCaenorhabditis elegansis an attractive model for studies of meiotic sister chromatid cohesion due to its genetic tractability and the excellent cytological properties of the hermaphrodite gonad. Moreover, mutants defective in the establishment or maintenance of meiotic SCC nevertheless produce abundant gametes, allowing analysis of the pattern of chromosome segregation. Here I describe two approaches for analysis of meiotic cohesion inC. elegans.The first approach relies on cytology to detect and quantify defects in SCC. The second approach relies on PCR and restriction digests to identify embryos that inherited an incorrect complement of chromosomes due to aberrant meiotic chromosome segregation. Both approaches are sensitive enough to identify rare errors and precise enough to reveal distinctive phenotypes resulting from mutations that perturb meiotic SCC in different ways. The robust, quantitative nature of these assays should strengthen phenotypic comparisons of different meiotic mutants and enhance the reproducibility of data generated by different investigators.