Bartnik E, Osborn M, Weber K
J Cell Biol. 1986 Jun;102(6):2033-41. doi: 10.1083/jcb.102.6.2033.
Current concepts of the developmentally controlled multigene family of intermediate filament (IF) proteins expect the origin of their complexity in evolutionary precursors preceding all vertebrate classes. Among invertebrates, however, firm ultrastructural as well as molecular documentation of IFs is restricted to some giant axons and to epithelia of a few molluscs and annelids. As Ascaris lumbricoides is easily dissected into clean tissues, IF expression in this large nematode was analyzed by electron microscopic and biochemical procedures and a monoclonal antibody reacting with all mammalian IF proteins. We document for the first time the presence of IFs in muscle cells of an invertebrate. They occur in three muscle types (irregular striated pharynx muscle, obliquely striated body muscle, uterus smooth muscle). IFs are also found in the epithelia studied (syncytial epidermis, intestine, ovary, testis). Immunoblots on muscles, pharynx, intestine, uterus, and epidermis identify a pair of polypeptides (with apparent molecular masses of 71 and 63 kD) as IF constituents. In vitro reconstitution of filaments was obtained with the proteins purified from body muscle. In the small nematode Caenorhabditis elegans IF proteins are so far found only in the massive desmosome-anchored tonofilament bundles which traverse a special epithelial cell type, the marginal cells of the pharynx. We speculate that IFs may occur in most but perhaps not all invertebrates and that they may not occur in all cells in large amounts. As electron micrographs of the epidermis of a planarian--a member of the Platyhelminthes--reveal IFs, the evolutionary origin of this cytoplasmic structure can be expected either among the lowest metazoa or already in some unicellular eukaryotes.
目前关于中间丝(IF)蛋白发育调控多基因家族的概念认为,其复杂性起源于所有脊椎动物类群之前的进化前体。然而,在无脊椎动物中,IF的可靠超微结构和分子证据仅限于一些巨大轴突以及少数软体动物和环节动物的上皮组织。由于蛔虫易于解剖成干净的组织,因此通过电子显微镜、生化方法以及一种能与所有哺乳动物IF蛋白发生反应的单克隆抗体,对这种大型线虫中的IF表达进行了分析。我们首次证明了无脊椎动物肌肉细胞中存在IF。它们存在于三种肌肉类型中(不规则横纹咽肌、斜纹体肌、子宫平滑肌)。在研究的上皮组织(合胞体表皮、肠道、卵巢、睾丸)中也发现了IF。对肌肉、咽、肠、子宫和表皮进行免疫印迹分析,确定了一对多肽(表观分子量分别为71 kD和63 kD)为IF成分。用从体肌中纯化的蛋白质实现了丝的体外重组。在小型线虫秀丽隐杆线虫中,迄今为止仅在大量的桥粒锚定张力丝束中发现IF蛋白,这些丝束穿过一种特殊的上皮细胞类型,即咽的边缘细胞。我们推测IF可能存在于大多数但也许不是所有的无脊椎动物中,并且它们可能并非在所有细胞中大量存在。由于涡虫(扁形动物门的一员)表皮的电子显微照片显示了IF,这种细胞质结构的进化起源可能在最低等的后生动物中,或者已经存在于一些单细胞真核生物中。