Institute of Molecular and Cellular Anatomy, RWTH Aachen University, Aachen, Germany.
School of Biological and Biomedical Sciences, Durham University, Durham, United Kingdom.
Sci Rep. 2020 Feb 21;10(1):3142. doi: 10.1038/s41598-020-59791-w.
The abundance and diversity of intermediate filaments (IFs) in the C. elegans intestine indicate important contributions to intestinal function and organismal wellbeing. Fluorescent IF reporters localize below the actin-rich brush border and are highly enriched in the lumen-enveloping endotube, which is attached to the C. elegans apical junction. Mapping intestinal viscoelasticity by contact-free Brillouin microscopy reveals that the IF-rich endotube is positioned at the interface between the stiff brush border and soft cytoplasm suggesting a mechanical buffering function to deal with the frequent luminal distortions occurring during food intake and movement. In accordance, depletion of IFB-2, IFC-2 and IFD-2 leads to intestinal lumen dilation although depletion of IFC-1, IFD-1 and IFP-1 do not. Ultrastructural analyses of loss of function mutants further show that IFC-2 mutants have a rarefied endotube and IFB-2 mutants lack an endotube altogether. Remarkably, almost all IFB-2- and IFC-2-deficient animals develop to fertile adults. But developmental retardation, reduced brood size, altered survival and increased sensitivity to microbial toxin, osmotic and oxidative stress are seen in both mutants albeit to different degrees. Taken together, we propose that individual intestinal IF polypeptides contribute in different ways to endotube morphogenesis and cooperate to cope with changing environments.
秀丽隐杆线虫肠道中中间丝(IFs)的丰富度和多样性表明它们对肠道功能和生物体健康有重要贡献。荧光 IF 报告蛋白定位于富含肌动蛋白的刷状缘下方,并且在包裹肠腔的内管中高度富集,内管附着在秀丽隐杆线虫的顶端连接上。通过无接触布里渊显微镜对肠道粘弹性进行映射,发现富含 IF 的内管位于坚硬的刷状缘和柔软的细胞质之间的界面处,这表明它具有机械缓冲功能,可以应对在进食和运动过程中经常发生的肠腔扭曲。相应地,尽管耗尽 IFC-1、IFD-1 和 IFP-1 不会导致这种情况,但 IFB-2、IFC-2 和 IFD-2 的耗竭会导致肠腔扩张。功能丧失突变体的超微结构分析进一步表明,IFC-2 突变体的内管稀少,而 IFB-2 突变体完全没有内管。值得注意的是,几乎所有 IFB-2 和 IFC-2 缺陷型动物都能发育为有生育能力的成虫。但是,在这两种突变体中都观察到了发育迟缓、繁殖力降低、存活率降低以及对微生物毒素、渗透压和氧化应激的敏感性增加,尽管程度不同。总之,我们提出,单个肠道 IF 多肽以不同的方式促进内管形态发生,并共同应对不断变化的环境。