Potenza Mariana, Tellez-Iñón María Teresa
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. H.N. Torres", INGEBI-CONICET, Vuelta de Obligado 2490, C1428ADN, Buenos Aires, Argentina,
Parasitol Res. 2015 Feb;114(2):641-9. doi: 10.1007/s00436-014-4227-8. Epub 2014 Nov 20.
This work analyzes the effect of the alkaloid colchicine on the growth of Trypanosoma cruzi epimastigotes, using immunofluorescence microscopy and flow cytometry techniques. We found that colchicine reversibly inhibited cytokinesis but not synthesis or segregation of nuclear and kinetoplastid DNA, in a concentration-dependent manner. We showed that, once colchicine was removed from the growth medium, cytokinesis was restored but abnormal segregation of kinetoplasts and nuclei generated zoids and parasites with two nuclei and one kinetoplast, among other aberrant cells. After drug removal, we also observed a few anucleated cells carrying two kinetoplasts in a stage compatible with the end of cytokinesis. The anomalous subcellular localization of the kinetoplast and flagellum observed in treated parasites suggests that the effect of colchicine and its interaction with T. cruzi microtubules is cell cycle dependent. The crosstalk between nuclear and kinetoplastid mitosis and its incidence on flagellum growth and parasite cell division regulation are discussed.
本研究利用免疫荧光显微镜和流式细胞术技术,分析了生物碱秋水仙碱对克氏锥虫前鞭毛体生长的影响。我们发现,秋水仙碱以浓度依赖的方式可逆地抑制胞质分裂,但不影响核DNA和动基体DNA的合成或分离。我们还发现,一旦从生长培养基中去除秋水仙碱,胞质分裂就会恢复,但动基体和细胞核的异常分离会产生具双鞭毛体和双细胞核单动基体的虫体以及其他异常细胞。去除药物后,我们还观察到一些无核细胞带有两个动基体,处于与胞质分裂末期相符的阶段。在经处理的虫体中观察到的动基体和鞭毛异常亚细胞定位表明,秋水仙碱的作用及其与克氏锥虫微管的相互作用是细胞周期依赖性的。本文还讨论了核有丝分裂与动基体有丝分裂之间的相互作用及其对鞭毛生长和寄生虫细胞分裂调控的影响。