Benchimol Marlene
Laboratòrio de Ultraestrutura Celular, Universidade Santa Ursula, Rua Jornalista Orlando Dantas, 59, CEP 222-31-010, Botafogo, Rio de Janeiro, Brazil.
Arch Microbiol. 2005 Mar;183(3):160-8. doi: 10.1007/s00203-004-0751-8. Epub 2005 Feb 10.
Giardia lamblia is a parasite possessing a complex cytoskeleton and an unusual morphology of bearing two nuclei. Here, the interphasic nuclei of trophozoites, using field emission scanning electron microscopy, routine scanning and transmission electron microscopy, immunocytochemistry, and 3D reconstruction, are presented. An approach using plasma-membrane extraction allowed the observation of the two nuclei still attached in their original positions. The observations are as follows: (1) Giardia nuclei and cytoskeleton were studied in demembranated cells by routine scanning electron microscopy and field emission; (2) both nuclei are anchored to basal bodies of the anterior flagella and to the descending posterior-lateral and ventral flagella, at the right and left nuclei, respectively, in cells attached by its ventral disc; (3) this attachment occurs by proteinaceous links, which were labeled by anti-actin and anti-centrin but not by anti-dynein or anti-tubulin antibodies; (4) fibrilar connections between the nuclei and the disc were also observed; and (5) nuclei exhibited a pendular movement when living cells were treated with cytochalasin, although the nuclei were still connected by their anterior region. Our analysis indicated that the nuclei have a defined position, and fibrils perform an anchoring system. This raises the possibility of a mechanism for nuclei-fidelity migration during mitosis.
蓝氏贾第鞭毛虫是一种具有复杂细胞骨架和独特形态(有两个细胞核)的寄生虫。本文利用场发射扫描电子显微镜、常规扫描和透射电子显微镜、免疫细胞化学以及三维重建技术,展示了滋养体间期细胞核的情况。一种利用质膜提取的方法使得我们能够观察到两个仍处于原始位置相连的细胞核。观察结果如下:(1)通过常规扫描电子显微镜和场发射技术,在去膜细胞中研究了蓝氏贾第鞭毛虫的细胞核和细胞骨架;(2)在通过腹吸盘附着的细胞中,两个细胞核分别通过蛋白质连接锚定在前鞭毛的基体以及右、左细胞核处向后下方延伸的后侧鞭毛和腹侧鞭毛上;(3)这种附着是通过蛋白质连接实现的,这些连接被抗肌动蛋白和抗中心蛋白抗体标记,但未被抗动力蛋白或抗微管蛋白抗体标记;(4)还观察到了细胞核与吸盘之间的纤维连接;(5)当用细胞松弛素处理活细胞时,细胞核表现出摆动运动,尽管细胞核仍通过其前部区域相连。我们的分析表明,细胞核具有确定的位置,并且纤维形成了一个锚定系统。这增加了在有丝分裂期间细胞核忠实迁移机制存在的可能性。