细胞技术

The kidney is a highly heterogeneous organ that is responsible for fluid and electrolyte balance. Much interest is focused on determining the function of specific renal epithelial cells in humans, which can only be accomplished through the isolation and growth of nephron segment-speci ...

The female reproductive system is a complex system. Epithelia of the female reproductive system including the ovaries, the oviduct, and the uterus are important sites for follicular development, ovulation, fertilization, implantation, and embryo development. They are also able to s ...

Most of the human aneuploidies have a maternal origin. This feature makes the study of human female meiosis a fundamental topic to understand the reasons leading to this important social problem. Unfortunately, due to sample collection difficulties, not many studies have been performed on ...

The basidiomycete fungus Coprinus cinereus has naturally synchronous meiosis and is amenable to analysis using an array of well-developed genetic and molecular tools. In this chapter, we explain in detail the two methods most commonly employed for C. cinereus, staining of intact gill seg ...

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 ...

A complex meiotic differentiation program generates genetically diverse haploid cells (gametes or spores) to compensate for the genome doubling that occurs at fertilization. To this end, homologous chromosomes must undergo pairing and recombination before they become parti ...

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 ...