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有丝分裂阻断微核畸变分析。

Cytokinesis Blocked Micronuclei Aberration Analysis.

机构信息

Department of Environmental & Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA.

出版信息

Methods Mol Biol. 2023;2519:83-91. doi: 10.1007/978-1-0716-2433-3_9.

Abstract

Cytokinesis blocked micronuclei (CBMN) assay is a rapid and sensitive analysis of chromosome aberrations and miss assortments during cell division. Genotoxic agent exposure produces DNA damage and chromosome fragments. Fragmented chromosomes without centromere failed to attach kinetochore which segregates a pair of homologous chromosomes to each daughter cells at cytokinesis, hence leading to form micronuclei. Chromosome or fragments of chromosome can also form micronuclei when they are not accurately sorted to daughter cells. Using cytochalasin B, an actin inhibitor, blocks cytokinesis of which completion leads serration appearance formed with two daughter cells while nuclei segregation is undergoing. As a result, one cell having two daughter nuclei, i.e., binucleated cell, is produced. By analyzing these binucleated cells, chromosome aberrations can be estimated as well as popular chromosome aberration analysis. Frequency of micronuclei formation predicts the testing agents' genotoxicity. By combining use with centromere-specific probes or DNA damage signal probes, the nature of genotoxicity of tested agents can be estimated.

摘要

细胞分裂阻断微核(CBMN)试验是一种快速而敏感的分析方法,可在细胞分裂过程中分析染色体畸变和错配。遗传毒性剂暴露会产生 DNA 损伤和染色体片段。没有着丝粒的片段化染色体无法附着动粒,而动粒将一对同源染色体在细胞分裂时分离到每个子细胞中,从而导致形成微核。当染色体或染色体片段不能准确地分配到子细胞中时,它们也会形成微核。细胞松弛素 B 是一种肌动蛋白抑制剂,可阻断细胞分裂,细胞分裂完成后会出现锯齿状外观,此时两个子细胞正在进行核分离。结果,一个细胞产生两个子核,即双核细胞。通过分析这些双核细胞,可以估计染色体畸变以及常见的染色体畸变分析。微核形成频率可预测测试剂的遗传毒性。通过与着丝粒特异性探针或 DNA 损伤信号探针结合使用,可以估计测试剂的遗传毒性性质。

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