Ribeiro Karla Consort, Mariante Rafael Meyer, Coutinho Letícia Lopes, Benchimol Marlene
Programa de Pós-Graduação em Ciências Morfológicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Biol Cell. 2002 Sep;94(4-5):289-301. doi: 10.1016/s0248-4900(02)01206-6.
We present observations on the fine structure and the division process of the nucleus in the protist Tritrichomonas foetus, parasite of the urogenital tract of cattle. The nucleus was followed by immunofluorescence and electron microscopy during interphase and mitosis. Conventional karyotyping coupled to image processing and bright field Panotic staining were used to follow nucleus modifications, chromosome number and condensation pattern along the whole cell cycle. Confocal laser scanning microscopy (CLSM) using DNA fluorescent probes, followed by image processing in the SURF-Driver program, produced three-dimensional reconstruction data of the mitotic nucleus under each phase of the division process. Immunocytochemistry in thin-sections revealed the chromosome spatial arrangement after bromodeoxyuridine incorporation and immunogold labeling using anti-DNA monoclonal antibodies. Our results indicate that: (1) the nucleus assumes different size and shapes along mitosis: it appears oval in interphase, becoming lobed or concave in prophase, then undergoing torsion and constriction, displaying an 'S' shape (metaphase). Next, it becomes elongated and it is finally separated in two nuclei at the transition of anaphase to telophase; (2) T. foetus nucleus harbors five chromosomes; (3) chromosomes become condensed in a pre-mitotic phase; (4) the nucleolus persists during the mitosis.
我们展示了对胎儿三毛滴虫(牛泌尿生殖道寄生虫)细胞核精细结构及分裂过程的观察结果。在间期和有丝分裂期间,通过免疫荧光和电子显微镜对细胞核进行追踪。使用传统核型分析结合图像处理和明场全染法来追踪整个细胞周期中细胞核的变化、染色体数目和凝聚模式。利用DNA荧光探针的共聚焦激光扫描显微镜(CLSM),随后在SURF-Driver程序中进行图像处理,生成了分裂过程各阶段有丝分裂细胞核的三维重建数据。薄切片免疫细胞化学揭示了溴脱氧尿苷掺入后染色体的空间排列以及使用抗DNA单克隆抗体的免疫金标记。我们的结果表明:(1)细胞核在有丝分裂过程中呈现不同的大小和形状:间期呈椭圆形,前期变为叶状或凹陷,随后发生扭转和收缩,呈现“S”形(中期)。接着,它变长,最终在后期向末期过渡时分裂为两个细胞核;(2)胎儿三毛滴虫细胞核含有五条染色体;(3)染色体在有丝分裂前期开始凝聚;(4)核仁在有丝分裂期间持续存在。