Aubusson-Fleury Anne, Cohen Jean, Lemullois Michel
Centre de Génétique Moléculaire, Institute for Integrative Biology of the Cell (I2BC), Université Paris Saclay, CEA, CNRS, Université Paris Sud, Bat 26 Allée de la terrasse, 91 198 Gif sur Yvette Cedex, France.
Methods Cell Biol. 2015;127:457-85. doi: 10.1016/bs.mcb.2014.12.007. Epub 2015 Feb 14.
Paramecium is a single cell able to divide in its morphologically differentiated stage that has many cilia anchored at its cell surface. Many thousands of cilia are thus assembled in a short period of time during division to duplicate the cell pattern while the cell continues swimming. Most, but not all, of these sensory cilia are motile and involved in two main functions: prey capture and cell locomotion. These cilia display heterogeneity, both in their length and their biochemical properties. Thanks to these properties, as well as to the availability of many postgenomic tools and the possibility to follow the regrowth of cilia after deciliation, Paramecium offers a nice opportunity to study the assembly of the cilia, as well as the genesis of their diversity within a single cell. In this paper, after a brief survey of Paramecium morphology and cilia properties, we describe the tools and the protocols currently used for immunofluorescence, transmission electron microscopy, and ultrastructural immunocytochemistry to analyze cilia, with special recommendations to overcome the problem raised by cilium diversity.
草履虫是一种单细胞生物，在其形态分化阶段能够进行分裂，其细胞表面锚定着许多纤毛。因此，在分裂过程中，数千根纤毛会在短时间内组装起来，以便在细胞继续游动的同时复制细胞模式。这些感觉纤毛中的大多数（但不是全部）是可运动的，并且参与两个主要功能：捕食和细胞运动。这些纤毛在长度和生化特性方面都表现出异质性。由于这些特性，以及许多后基因组工具的可用性和去纤毛后追踪纤毛再生的可能性，草履虫为研究纤毛的组装以及单个细胞内纤毛多样性的起源提供了一个很好的机会。在本文中，在简要概述草履虫的形态和纤毛特性之后，我们描述了目前用于免疫荧光、透射电子显微镜和超微结构免疫细胞化学分析纤毛的工具和方案，并特别提出了克服纤毛多样性所带来问题的建议。
