Lefort-Tran M, Aufderheide K, Pouphile M, Rossignol M, Beisson J
J Cell Biol. 1981 Feb;88(2):301-11. doi: 10.1083/jcb.88.2.301.
The trichocysts of Paramecium tetraurelia constitute a favorable system for studying secretory process because of the numerous available mutations that block, at various stages, the development of these secretory vesicles, their migration towards and interaction with the cell surface, and their exocytosis. Previous studies of several mutants provided information (a) on the assembly and function of the intramembranous particles arrays in the plasma membrane at trichocyst attachment sites, (b) on the autonomous motility of trichocysts, required for attachment to the cortex, and (c) on a diffusible cytoplasmic factor whose interaction with both trichocyst and plasma membrane is required for exocytosis to take place. We describe here the properties of four more mutants deficient in exocytosis ability, nd6, nd7, tam38, and tam6, which were analyzed by freeze-fracture, microinjection of trichocysts, and assay for repair of the mutational defect through cell-cell interaction during conjugation with wild-type cells. As well as providing confirmation of previous conclusions, our observations show that the mutations nd6 and tam6 (which display striking abnormalities in their plasma membrane particle arrays and are reparable through cell-cell contact but not by microinjection of cytoplasm) affect two distinct properties of the plasma membrane, whereas the other two mutations affect different properties of the trichocysts. Altogether, the mutants so far analyzed now provide a rather comprehensive view of the steps and functions involved in secretory processes in Paramecium and demonstrate that two steps of these processes, trichocyst attachment to the plasma membrane and exocytosis, depend upon specific properties of both the secretory vesicle and the plasma membrane.
由于有大量可用的突变体,它们在不同阶段阻断了这些分泌小泡的发育、它们向细胞表面的迁移和与细胞表面的相互作用以及它们的胞吐作用,四膜虫的刺丝泡构成了一个研究分泌过程的良好系统。先前对几个突变体的研究提供了以下信息:(a) 关于刺丝泡附着位点处质膜中膜内颗粒阵列的组装和功能;(b) 关于刺丝泡附着到皮层所需的自主运动性;(c) 关于一种可扩散的细胞质因子,胞吐作用发生需要它与刺丝泡和质膜都相互作用。我们在此描述另外四个胞吐能力缺陷的突变体nd6、nd7、tam38和tam6的特性,通过冷冻断裂、刺丝泡显微注射以及在与野生型细胞接合期间通过细胞 - 细胞相互作用修复突变缺陷的测定来分析这些突变体。除了证实先前的结论外,我们的观察结果表明,nd6和tam6突变(它们在质膜颗粒阵列中表现出明显异常,可通过细胞 - 细胞接触修复,但不能通过细胞质显微注射修复)影响质膜的两个不同特性,而另外两个突变影响刺丝泡的不同特性。总之,到目前为止分析的这些突变体现在提供了四膜虫分泌过程中所涉及步骤和功能的相当全面的观点,并证明这些过程的两个步骤,即刺丝泡附着到质膜和胞吐作用,取决于分泌小泡和质膜的特定特性。