细胞功能测定

The filamentous fungi Neurospora crassa and Sordaria macrospora are materials of choice for recombination studies because each of the DNA strands involved in meiosis can be visually analyzed using spore-color mutants. Well-advanced molecular genetic methodologies have been d ...

Important information on cellular physiology can be obtained by directly observing living cells. The nucleus, and the chromatin within, is of particular interest to many researchers. Monitoring the behavior of specific DNA loci in the living cell is now commonly achieved through the inse ...

Movements are implicit in the chromosome behaviors of bouquet formation, pairing and synapsis during meiotic prophase. In S. cerevisiae, the positions of chromosomes, specific structures, and individual chromosomal loci marked by fluorescent fusion proteins are easily visual ...

The fission yeast Schizosaccharomyces pombe has provided a useful experimental system to study nuclear structures during meiosis. Unlike many higher animals in which meiosis takes place only in specialized tissues deep inside their bodies, S. pombe is a unicellular eukaryote and its m ...

Fluorescent in situ hybridization (FISH) provides a powerful tool to study the localization of DNA sequences in relationship to one another. FISH has the advantage over other methods, notably use of GFP-tagged repressor/operator arrays, that an almost unlimited number of probes can be uti ...

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 pr ...

Visualization of meiotic chromosomes in the model organism S. cerevisiae has become an integral part of the study of wild-type meiosis and the characterization of mutant phenotypes. This chapter describes a simple method for chromosome spreading, which is a variation on a protocol origin ...

Advances in molecular biology and in the genetics of Arabidopsis thaliana have led to this organism becoming an important model for the analysis of meiosis in plants. Cytogenetic investigations are pivotal to meiotic studies and a number of technological improvements for Arabidopsis ...

In recent years, zebrafish (Danio rerio) has been used as a model vertebrate organism for studies of human disease, development, and genetics. This chapter describes detailed methods for the preparation of whole-mount meiotic oocytes and spermatocytes as well as cryostat sectioning of o ...

The genus Daphnia has an intriguing reproductive mode of cyclical parthenogenesis. This reproductive mode has been studied for centuries, but cytogenetic information is lacking due to technical limitations of classical methods. We have developed methods for the preparation and e ...

A wide variety of techniques have been utilized to determine the localization of various proteins from premeiotic through meiotic stages in Drosophila males. Live imaging has been instrumental in monitoring chromosome pairing and the localization of fusion proteins. Immunofluo ...

Methods are described to analyze two different parts of the Drosophila ovary, which correspond to early stages (pachytene) and late stages (metaphase I and beyond) of meiosis. In addition to taking into account morphology, the techniques differ by fixation conditions and the method to isol ...

The nematode Caenorhabditis elegans has emerged as an informative experimental system for analysis of meiosis, in large part because of the advantageous physical organization of meiotic nuclei as a gradient of stages within the germline. Here we provide tools for detailed observatio ...

Many of the structures involved in meiotic synapsis and recombination such as synaptonemal complexes (SCs) and recombination nodules (RNs) can be resolved only by electron microscopy. Therefore, electron microscopic (EM) immunolocalization using gold-conjugated antibod ...

Spermatogenesis is a cyclic process during which, within each epithelial area, various generations of germ cells undergo a series of developmental steps according to a fixed time schedule. The cycle of the seminiferous epithelium can be subdivided into stages. In the mouse, 12 such stages ha ...

Understanding meiosis is facilitated by in vitro experimental approaches, but this has not been easily applicable to mammalian meiocytes. Available methods for in vitro analysis of mammalian oocytes are generally limited to experimental analysis of the late prophase period. Short ...

The studies of molecular events that occur in single cell types within a tissue often require the disaggregation of the tissue into a single cell suspension, followed by isolation of distinct cell populations. The germinal epithelium of mammals is composed of several cell types, which divide ...

The goal of this chapter is twofold: First, to acquaint the reader with the problems inherent in analyzing mammalian female meiosis and, second, to provide a step-by-step approach to mastering the necessary techniques. Although the methods presented are for use in the human and mouse, with subtle ...

New immunofluorescence techniques allow visual identification of human cells in various stages of meiotic prophase. Antibodies to the synaptonemal complex, the centromere and sites of recombination allow these stages of meiotic prophase to be identified. The progress of chromo ...

During mouse meiosis, gene expression and homologous synapsis are intimately linked. Chromosomes that fail to synapse at the zygotene–pachytene transition become transcriptionally silenced by a process called Meiotic Silencing of Unsynapsed Chromatin (MSUC), and this sile ...