Cytological Analysis of Interference in Mouse Meiosis
Cytological Analysis of Interference in Mouse Meiosis
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DOI：
10.1007/978-1-60761-103-5_21
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发表时间：
2009-01-01
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影响因子：
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通讯作者：
Heyting, Christa
中科院分区：
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作者：
de Boer, Esther;Lhuissier, Franck G. P.;Heyting, Christa
在大多数真核生物中，减数分裂交叉 (CO) 沿着二价体非随机放置，因此 CO 的存在降低了附近出现其他 CO 的可能性。这种现象被称为 CO 干扰，最初是在遗传学上定义的，但也可以通过研究参与 CO 形成的蛋白质复合物的染色体位置或通过研究交叉的位置来进行细胞学分析。在这里，我们重点关注参与小鼠减数分裂重组和 CO 形成的蛋白质复合物之间的干扰的细胞学分析。在减数分裂的粗线期，这些蛋白质复合物可以被可视化为沿着联会复合物（SC）的免疫荧光焦点，联会复合物是在减数分裂前期沿着同源染色体对（二价体）形成的线性蛋白质结构。我们描述了如何制作适合分析这些焦点之间的干扰的细胞学制剂，以及如何使用伽马分布作为焦点/CO 定位的数学模型来估计焦点之间的干扰强度。
In most eukaryotes, meiotic crossovers (COs) are non-randomly placed along the bivalents, such that the presence of a CO reduces the probability of additional COs nearby. This phenomenon, named CO interference, was originally defined genetically, but can also be analyzed cytologically by studying the chromosomal positions of protein complexes that are involved in CO formation, or by studying the positions of chiasmata. Here we focus on the cytological analysis of interference among protein complexes involved in meiotic recombination and CO formation in the mouse. During the pachytene stage of meiosis, these protein complexes can be visualized as immunofluorescent foci along synaptonemal complexes (SCs), which are linear protein structures that are formed along homologous chromosome pairs (bivalents) during meiotic prophase. We describe how to make cytological preparations that are suitable for the analysis of interference among these foci, and how to estimate the strength of interference among foci, using the gamma distribution as a mathematical model for focus/CO positioning.