Department of Cell Biology and Genetics, Erasmus MC, Rotterdam, the Netherlands.
J Cell Sci. 2010 Jun 15;123(Pt 12):2077-84. doi: 10.1242/jcs.062885. Epub 2010 May 25.
Cilia length and function are dynamically regulated by modulation of intraflagellar transport (IFT). The cilia of C. elegans amphid channel neurons provide an excellent model to study this process, since they use two different kinesins for anterograde transport: kinesin-II and OSM-3 kinesin together in the cilia middle segments, but only OSM-3 in the distal segments. To address whether sensory signaling modulates the coordination of the kinesins, we studied IFT protein motility in gpa-3 mutant animals, since dominant active mutation of this sensory Galpha protein GPA-3QL) affects cilia length. In addition, we examined animals exposed to dauer pheromone, since dauer formation, which involves gpa-3, induces changes in cilia morphology. Live imaging of fluorescently tagged IFT proteins showed that in gpa-3 mutants and in larvae exposed to dauer pheromone, kinesin-II speed is decreased and OSM-3 speed is increased, whereas structural IFT proteins move at an intermediate speed. These results indicate that mutation of gpa-3 and exposure to dauer pheromone partially uncouple the two kinesins. We propose a model in which GPA-3-regulated docking of kinesin-II and/or OSM-3 determines entry of IFT particles into the cilia subdomains, allowing structural and functional plasticity of cilia in response to environmental cues.
纤毛长度和功能通过鞭毛内运输(IFT)的调节而动态变化。秀丽隐杆线虫触角通道神经元的纤毛为研究这一过程提供了极好的模型,因为它们使用两种不同的驱动蛋白进行正向运输:中间纤毛段使用驱动蛋白-II 和 OSM-3 驱动蛋白,而在远段仅使用 OSM-3 驱动蛋白。为了研究感觉信号是否调节驱动蛋白的协调,我们研究了 gpa-3 突变体动物中的 IFT 蛋白运动,因为该感觉 G 蛋白 GPA-3QL 的显性激活突变会影响纤毛长度。此外,我们还检查了暴露于 dauer 信息素的动物,因为 dauer 形成(涉及 gpa-3)会导致纤毛形态发生变化。荧光标记的 IFT 蛋白的实时成像显示,在 gpa-3 突变体和暴露于 dauer 信息素的幼虫中,驱动蛋白-II 的速度降低,OSM-3 的速度增加,而结构 IFT 蛋白以中间速度移动。这些结果表明,gpa-3 的突变和 dauer 信息素的暴露部分使两种驱动蛋白解偶联。我们提出了一个模型,即 GPA-3 调节的驱动蛋白-II 和/或 OSM-3 的对接决定了 IFT 颗粒进入纤毛亚域的入口,从而允许纤毛对环境信号做出结构和功能的可塑性反应。