Use of Ciliogenesis to Detect Aneugens: The Role of Primary Cilia

Kathyayini V. Divi, Yvona Ward, Miriam C. Poirier, Ofelia A. Olivero
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引用次数: 1

Abstract

Primary cilia arise from the centrosomes of quiescent or post-mitotic cells, and serve as sensory organelles that communicate mechanical and chemical stimuli from the environment to the interior of the cell. Cilium formation may, therefore, become a useful end point signaling exposure to genotoxins or aneugens. Here we have used the aneugen, zidovudine (AZT), an antiretroviral drug that induces DNA replication arrest and centrosomal amplification (>2 centrosomes per quiescent cell), to evaluate cilia formation in retinal epithelial (pigmented) cells. Since cilia are derived from centrosomes, and aneugens can induce centrosomal amplification, the production of multiple cilia arising from multiple centrosomes may reveal the aneugenic nature of the agents. Cells were exposed to AZT to induce centrosomal amplification, cultured without serum to allow the centrioles to develop cilia, and immunostained to visualize cilia and centrosomes. Nuclear DNA was stained with DAPI. Preliminary observations suggest that cells with multiple centrosomes are able to generate extra cilia. © 2015 by John Wiley & Sons, Inc.

用纤毛发生检测纤毛:初级纤毛的作用
初级纤毛起源于静止细胞或有丝分裂后细胞的中心体,作为感觉细胞器,将来自环境的机械和化学刺激传递到细胞内部。因此,纤毛的形成可能成为基因毒素或基因原暴露的有用终点信号。在这里,我们使用齐多夫定(AZT),一种诱导DNA复制阻滞和中心体扩增(每个静止细胞2个中心体)的抗逆转录病毒药物,来评估视网膜上皮(色素)细胞的纤毛形成。由于纤毛来源于中心体,而非优生原可以诱导中心体扩增,因此由多个中心体产生多个纤毛可能揭示了药物的非优生性质。细胞暴露于AZT诱导中心体扩增,无血清培养使中心粒发育纤毛,免疫染色观察纤毛和中心体。DAPI染色细胞核DNA。初步观察表明,具有多个中心体的细胞能够产生额外的纤毛。©2015 by John Wiley &儿子,Inc。
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