Loidl Josef, Lorenz Alexander
Department of Chromosome Biology, Center for Molecular Biology, University of Vienna, Vienna, Austria.
Methods Mol Biol. 2009;558:15-36. doi: 10.1007/978-1-60761-103-5_2.
The fission yeast, Schizosaccharomyces pombe, much like the budding yeast, is a particularly well-suited model organism for genetic research. However, the miniscule size of both yeasts' nuclei has hindered their success as research models for cytologists. A solution to this problem is provided by the spreading of nuclei, which increases their volume and allows for a better spatial resolution of nuclear contents. Here we describe nuclear spreading in fission yeast. Spreading of meiotic nuclei is particularly helpful in exposing the linear elements (LinEs), which are the fission yeasts' rudimentary version of the synaptonemal complex. Although the LinEs' role is still not fully understood, they serve as important meiotic hallmarks and their presence and morphology can be used in characterizing meiotic mutants. We first describe methods to induce meiosis in liquid cell cultures, then outline a method to break down cell and nuclear membranes by detergent treatment to release chromatin on cytological slides, and finally provide a set of protocols for analyzing these nuclei by immunostaining and fluorescence in situ hybridization (FISH), and by electron microscopy.
裂殖酵母，即粟酒裂殖酵母，与芽殖酵母非常相似，是一种特别适合用于基因研究的模式生物。然而，这两种酵母细胞核的微小尺寸阻碍了它们成为细胞学家研究的成功模型。细胞核铺展为这个问题提供了解决方案，它增加了细胞核的体积，并能更好地对核内容物进行空间分辨。在此，我们描述裂殖酵母中的细胞核铺展。减数分裂细胞核的铺展对于揭示线性元件（LinEs）特别有帮助，这些线性元件是裂殖酵母中突触复合体的初级形式。尽管线性元件的作用仍未完全了解，但它们是重要的减数分裂标志，其存在和形态可用于鉴定减数分裂突变体。我们首先描述在液体细胞培养物中诱导减数分裂的方法，然后概述通过去污剂处理破坏细胞膜和核膜以在细胞学载玻片上释放染色质的方法，最后提供一套通过免疫染色和荧光原位杂交（FISH）以及电子显微镜分析这些细胞核的方案。
