Carvalho Karina Penedo, Monteiro-Leal Luiz Henrique
Laboratório de Microscopia e Processamento de Imagens, Departemento de Histologia e Embriologia, Universidade do Estado do Rio de Janeiro, Av. Prof. Manoel de Abreu, 444-3 andar, Maracanã Rio de Janeiro, RJ 20550-170, Brazil.
Exp Parasitol. 2004 Nov-Dec;108(3-4):154-62. doi: 10.1016/j.exppara.2004.08.007.
This paper presents a detailed study of the caudal complex of Giardia lamblia and its relation to movements observed in this region. The caudal complex of Giardia, composed of axonemes from the caudal flagella plus associated microtubular sheets, was investigated by light, electron microscopy, and 3D reconstruction tools. By the use of video-microscopy and digital image processing techniques, we were able to visualize in detail the caudal movements. A non-ionic detergent, Triton X-100, was used to isolate the complex that was afterwards analyzed by video-microscopy and transmission electron microscopy (TEM). We showed for the first time, using video-microscopy, that the intracellular portion of the caudal flagella axonemes presented motility, even after the disrupture of the cell membrane, contrasting with the caudal flagella themselves, that do not show active beating pattern. To check if actin filaments play a role in the above described movements, as previously supposed, we incubated the cells with jasplakinolide, a drug that induces the disruption of actin filaments in living cells. The experiments demonstrated that the drug did not affect the caudal motility. The analysis of the caudal complex by transmission electron microscopy (TEM) revealed that, even after the exposure to higher detergent concentrations, the connections between their components remained intact. The information obtained by TEM and 3D reconstruction tools showed that the region between both nuclei marks the intracellular end of the caudal complex, which proceeds toward the caudal portion of the cell following its longitudinal axis, where the axonemes emerge as the caudal flagella. The results obtained from video-microscopy assays of the isolated beating complex together with the 3D reconstruction data indicated that the internal portion of the caudal flagella is the force-generator of the movements in this region.
本文对蓝氏贾第鞭毛虫的尾复合体及其与该区域观察到的运动的关系进行了详细研究。通过光学显微镜、电子显微镜和三维重建工具,对由尾鞭毛轴丝及相关微管片组成的蓝氏贾第鞭毛虫尾复合体进行了研究。利用视频显微镜和数字图像处理技术,我们能够详细观察尾运动。使用非离子去污剂Triton X-100分离该复合体,随后通过视频显微镜和透射电子显微镜(TEM)进行分析。我们首次通过视频显微镜显示,即使在细胞膜破裂后,尾鞭毛轴丝的细胞内部分仍具有运动性,这与尾鞭毛本身不同,尾鞭毛不呈现主动摆动模式。为了检验肌动蛋白丝是否如先前推测的那样在上述运动中发挥作用,我们用茉莉素内酯处理细胞,茉莉素内酯是一种能诱导活细胞中肌动蛋白丝断裂的药物。实验表明该药物不影响尾运动。通过透射电子显微镜(TEM)对尾复合体的分析表明,即使在暴露于较高去污剂浓度后,其各组分之间的连接仍保持完整。通过TEM和三维重建工具获得的信息表明,两个细胞核之间的区域标志着尾复合体的细胞内末端,该末端沿着细胞的纵轴朝向细胞的尾部延伸,轴丝在尾部作为尾鞭毛出现。从分离的摆动复合体的视频显微镜检测结果以及三维重建数据表明,尾鞭毛的内部是该区域运动的力产生部位。