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Colchicine-induced polyploidization depends on tubulin polymerization in c-metaphase cells

机译:秋水仙碱诱导的多倍体化取决于c中期细胞中的微管蛋白聚合

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摘要

The microtubule cytoskeleton plays a crucial role in the cell cycle and in mitosis. Colchicine is a microtubule-depolymerizing agent that has long been used to induce chromosome individualization in cells arrested at metaphase and also in the induction of polyploid plants. Although attempts have been made to explain the processes and mechanisms underlying polyploidy induction, the role of the cytoskeleton still remains largely unknown. Through immunodetection of alpha-tubulin, different concentrations (0.5 or 5 mM) of colchicine were found to produce opposite effects in the organization of the cytoskeleton in rye (Secale cereale L.). A low concentration (0.5 mM) induced depolymerization of the microtubular cytoskeleton in all phases of the cell cycle. In contrast, a high concentration (5 mM) was found to induce the polymerization of new tubulin-containing structures in c-metaphase cells. Furthermore, both treatments also showed contrasting effects in the induction of polyploid cells. Flow cytometric analysis and quantitative assessments of nucleolus-organizing regions revealed that only the high-concentration colchicine treatment was effective in the formation of polyploid cells. Our studies indicate that spindle disruption alone is insufficient for the induction of polyploid cells. The absence of any tubulin structures in plants treated with colchicine at the low concentration induced cell anomalies, such as the occurrence of nuclei with irregular shape and/or (additional) micronuclei, 12 h after recovery, pointing to a direct effect on cell viability. In contrast, the almost insignificant level of cell anomalies in the high-concentration treatment suggests that the presence of new tubulin-containing structures allows the reconstitution of 4C nuclei and their progression into the cell cycle.
机译:微管细胞骨架在细胞周期和有丝分裂中起关键作用。秋水仙碱是一种微管解聚剂,长期以来一直用于诱导在中期停滞的细胞中的染色体个体化,以及在多倍体植物的诱导中。尽管已经尝试解释多倍体诱导的过程和机制,但是细胞骨架的作用仍然很大程度上未知。通过对α-微管蛋白的免疫检测,发现不同浓度(0.5或5 mM)的秋水仙碱在黑麦(Secale graine L.)的细胞骨架组织中产生相反的作用。低浓度(0.5 mM)在细胞周期的所有阶段均诱导微管细胞骨架解聚。相反,发现高浓度(5 mM)可以诱导c中期细胞中新的含微管蛋白的结构聚合。此外,两种处理在多倍体细胞的诱导中也显示出相反的作用。流式细胞仪分析和核仁组成区域的定量评估表明,只有高浓度秋水仙碱处理才能有效地形成多倍体细胞。我们的研究表明,仅纺锤体破坏不足以诱导多倍体细胞。在低浓度的秋水仙碱处理过的植物中,没有任何微管蛋白结构会诱导细胞异常,例如在恢复后12 h出现形状不规则和/或(附加)微核的核,这直接影响了细胞活力。相反,在高浓度处理中,细胞异常的水平几乎不显着,这表明新的含微管蛋白的结构的存在允许4C核的重建及其进入细胞周期的进程。

著录项

  • 来源
    《Protoplasma》 |2006年第4期|147-153|共7页
  • 作者单位

    Secção de Genética Centro de Botânica Aplicada à Agricultura Instituto Superior de Agronomia Universidade Técnica de LisboaDepartamento de Ciências Naturais Ambientais e Biotecnológicas Universidade Lusófona de Humanidades e Tecnologias;

    Secção de Genética Centro de Botânica Aplicada à Agricultura Instituto Superior de Agronomia Universidade Técnica de LisboaDepartamento de Ciências Naturais Ambientais e Biotecnológicas Universidade Lusófona de Humanidades e Tecnologias;

    Secção de Genética Centro de Botânica Aplicada à Agricultura Instituto Superior de Agronomia Universidade Técnica de Lisboa;

    Karyotype Evolution Group Institut für Pflanzengenetik und Kulturpflanzenforschung;

    Institute of Biological Sciences University of Wales;

    Secção de Genética Centro de Botânica Aplicada à Agricultura Instituto Superior de Agronomia Universidade Técnica de Lisboa;

    Chromosome Structure and Function Group Institut für Pflanzengenetik und Kulturpflanzenforschung;

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  • 原文格式 PDF
  • 正文语种 eng
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  • 关键词

    Keywords: Induced polyploidy; Colchicine; Tubulin array; Secale cereale.;

    机译:关键词:诱导多倍体秋水仙碱;小管蛋白阵列;麦片粥。;

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