School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang, 453003, China.
School of Pharmacy, Xinxiang Medical University, Xinxiang, 453003, China.
J Eukaryot Microbiol. 2020 Sep;67(5):521-531. doi: 10.1111/jeu.12799. Epub 2020 May 25.
Intraflagellar transport (IFT) represents a bidirectional dynamic process that carries cargo essential for cilia building and the maintenance of ciliary function, which is important for the locomotion of single cells, intracellular and intercellular signalling transduction. Accumulated evidence has revealed that defects in IFT cause several clinical disorders. Here, we determined the role of IFT80, an IFT-B protein that is mutated in Jeune asphyxiating thoracic dystrophy. Using the RNAi method in the ciliate Paramecium as model, we found that loss of IFT80 prevents cilia biogenesis and causes strong cell lethality. A specific antibody against IFT80 was also prepared in our study, which labelled IFT80 in cilia of Paramecium. GFP fusion experiments were performed to illustrate the dynamic movement of IFT-A and IFT-B proteins in cilia of Paramecium; then, we found that the depletion of IFT80 in cells prevents IFT-A and IFT-B proteins from entering the cilia. Our results showed the distribution change of other IFT proteins in cells that were depleted of IFT80, and we discuss the possible roles of IFT80 in Paramecium.
纤毛内运输（IFT）代表了一种双向动态过程，它携带对纤毛构建和纤毛功能维持至关重要的货物，这对于单细胞的运动、细胞内和细胞间信号转导很重要。越来越多的证据表明，IFT 的缺陷会导致几种临床疾病。在这里，我们确定了 IFT-B 蛋白 IFT80 的作用，IFT80 在 Jeune 窒息性胸肌营养不良中发生突变。我们使用纤毛生物 Paramecium 中的 RNAi 方法作为模型，发现 IFT80 的缺失会阻止纤毛发生，并导致强烈的细胞致死。在我们的研究中，还制备了针对 IFT80 的特异性抗体，该抗体标记了 Paramecium 纤毛中的 IFT80。进行 GFP 融合实验来阐明 Paramecium 纤毛中 IFT-A 和 IFT-B 蛋白的动态运动；然后，我们发现细胞中 IFT80 的耗竭会阻止 IFT-A 和 IFT-B 蛋白进入纤毛。我们的结果显示了细胞中耗尽 IFT80 后其他 IFT 蛋白的分布变化，并讨论了 IFT80 在 Paramecium 中的可能作用。
