Alcantara Carolina de L, Vidal Juliana C, de Souza Wanderley, Cunha-E-Silva Narcisa L
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, Brazil.
Núcleo de Biologia Estrutural e Bioimagens (CENABIO), Rio de Janeiro 21941-902, Brazil.
J Cell Sci. 2017 Jan 1;130(1):164-176. doi: 10.1242/jcs.187419. Epub 2016 Jun 30.
The cytostome-cytopharynx complex is the main site for endocytosis in epimastigotes of Trypanosoma cruzi It consists of an opening at the plasma membrane surface - the cytostome - followed by a membrane invagination - the cytopharynx. In G1/S cells, this structure is associated with two specific sets of microtubules, a quartet and a triplet. Here, we used electron microscopy and electron tomography to build 3D models of the complex at different stages of the cell cycle. The cytostome-cytopharynx is absent in late G2 and M phase cells, whereas early G2 cells have either a short cytopharynx or no visible complex, with numerous vesicles aligned to the cytostome-cytopharynx microtubules. The microtubule quartet remains visible throughout cell division (albeit in a shorter form), and is duplicated during G2/M. In contrast, the microtubule triplet is absent during late G2/M. Cells in cytokinesis have an invagination of the flagellar pocket membrane likely to represent early stages in cytostome-cytopharynx assembly. Cells in late cytokinesis have two fully developed cytostome-cytopharynx complexes. Our data suggest that the microtubule quartet serves as a guide for new cytostome-cytopharynx assembly.
胞口-胞咽复合体是克氏锥虫上鞭毛体中内吞作用的主要部位。它由质膜表面的一个开口——胞口,以及随后的一个膜内陷——胞咽组成。在G1/S期细胞中,这种结构与两组特定的微管相关联,一组是四重微管,另一组是三重微管。在这里,我们使用电子显微镜和电子断层扫描技术构建了细胞周期不同阶段该复合体的三维模型。在G2晚期和M期细胞中不存在胞口-胞咽,而G2早期细胞要么有一个短的胞咽,要么没有可见的复合体,但有许多囊泡与胞口-胞咽微管排列在一起。四重微管在整个细胞分裂过程中都可见(尽管形式较短),并在G2/M期复制。相比之下,三重微管在G2/M晚期不存在。处于胞质分裂期的细胞有鞭毛袋膜内陷,这可能代表胞口-胞咽组装的早期阶段。处于胞质分裂后期的细胞有两个完全发育的胞口-胞咽复合体。我们的数据表明,四重微管为新的胞口-胞咽组装提供了导向。