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药物诱导的早熟染色体凝聚(PCC)方案:有丝分裂染色体和间期染色质的细胞遗传学方法

Drug-induced premature chromosome condensation (PCC) protocols: cytogenetic approaches in mitotic chromosome and interphase chromatin.

作者信息

Gotoh Eisuke

机构信息

Division of Genetic Resources, National Institute of Infectious Diseases, Tokyo, Japan.

出版信息

Methods Mol Biol. 2009;523:83-92. doi: 10.1007/978-1-59745-190-1_6.

Abstract

Chromosome analysis is a fundamental technique for cytogenetic studies. Chromosomes are conventionally prepared from mitotic cells arrested by colcemid block protocol. However, obtaining the mitotic chromosomes is often hampered under several circumstances. As a result, cytogenetic analysis will be sometimes difficult or even impossible in such cases. Premature chromosome condensation (PCC) is an alternative method that has proved to be a unique and useful way in chromosome analysis. Usually, PCC has been achieved following cell fusion mediated either by fusogenic viruses or by polyethylene glycol (cell-fusion PCC), but the cell-fusion PCC has several drawbacks. The novel drug-induced PCC using protein phosphatase inhibitors was introduced about 10 years ago. This method is much simple and easy even than the conventional mitotic chromosome preparation using colcemid block protocol and obtained PCC index (equivalent to mitotic index for metaphase chromosome) is much higher. Furthermore, this method allows the interphase chromatin to be condensed and visualized like mitotic chromosomes, thus opened the way for chromosome analysis not only in metaphase chromosomes but also in interphase chromatin. The drug-induced PCC has therefore proven the usefulness in cytogenetics and other cell biology fields.

摘要

染色体分析是细胞遗传学研究的一项基本技术。传统上,染色体是从经秋水仙酰胺阻断方案阻滞的有丝分裂细胞中制备的。然而,在几种情况下,获取有丝分裂染色体常常受到阻碍。因此,在这种情况下,细胞遗传学分析有时会很困难甚至无法进行。早熟染色体凝集(PCC)是一种替代方法,已被证明是染色体分析中一种独特且有用的方式。通常,PCC是在由融合病毒或聚乙二醇介导的细胞融合后实现的(细胞融合PCC),但细胞融合PCC有几个缺点。大约10年前引入了使用蛋白磷酸酶抑制剂的新型药物诱导PCC。这种方法甚至比使用秋水仙酰胺阻断方案的传统有丝分裂染色体制备方法更简单易行,并且获得的PCC指数(相当于中期染色体的有丝分裂指数)要高得多。此外,这种方法可以使间期染色质像有丝分裂染色体一样凝集并可视化,从而不仅为中期染色体的分析,也为间期染色质的分析开辟了道路。因此,药物诱导PCC已证明在细胞遗传学和其他细胞生物学领域是有用的。

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