Phaire-Washington L, Howard-Robinson G, Lucas J, Roberson J, Brako E, McNeil N, Williams S, Watson C, Igbo T, Brako L
Scanning Microsc. 1987 Jun;1(2):775-82.
This study characterizes two- and three-dimensional ultrastructure and surface topography of polymerized networks of intermediate filaments (IF) isolated from mouse peritoneal macrophages. Isolated IF bound to monoclonal anti-IF antibodies in enzyme-linked immunosorbent assays. Immunogold labeling of IF with specific antibodies revealed that epitopes are distributed along filaments particularly at junctions where filaments interconnect. Networks of IF, viewed by scanning electron microscopy, organized as ropelike groups of interconnecting filaments which swirl and encircle each other to form three-dimensional lattices containing ellipsoidal-, circular-, and vacuole-shaped cavities. Cavity diameters were similar in size to organelles and vacuoles; diameters were grouped as small (12-288 nm), medium (0.3-1.7 micron), and large (2-3 micron). The walls of the cavities appeared as beaded structures with alternating globular and linear regions. Linear regions were 14 nm. Repeat distances taken from the central axis of globular regions were 23-27 nm. The lattice organization of IF observed in vitro was similar to images seen in vivo in Triton-insoluble cytoskeletons immunofluorescently labeled with specific antibodies. In whole cells processed for TEM, swirling bundles of IF were found encircling membranous vacuoles. Based on the lattice architecture of IF, cavity dimensions, and IF location, we postulate that intermediate filaments may function in the mechanical and spatial distribution of vacuoles in the cell cytoplasm.
本研究对从小鼠腹腔巨噬细胞中分离出的中间丝(IF)聚合网络的二维和三维超微结构及表面形貌进行了表征。在酶联免疫吸附测定中,分离出的IF与单克隆抗IF抗体结合。用特异性抗体对IF进行免疫金标记显示,表位沿细丝分布,尤其在细丝相互连接的连接处。通过扫描电子显微镜观察,IF网络组织成相互连接的细丝状绳状组,它们相互缠绕并环绕形成包含椭圆形、圆形和液泡形腔的三维晶格。腔的直径与细胞器和液泡的大小相似;直径分为小(12 - 288纳米)、中(0.3 - 1.7微米)和大(2 - 3微米)。腔壁呈现为具有交替球状和线性区域的串珠状结构。线性区域为14纳米。从球状区域中心轴测量的重复距离为23 - 27纳米。在体外观察到的IF晶格组织与在用特异性抗体进行免疫荧光标记的Triton不溶性细胞骨架中在体内看到的图像相似。在用于透射电子显微镜的完整细胞中,发现IF的缠绕束环绕着膜性液泡。基于IF的晶格结构、腔的尺寸和IF的位置,我们推测中间丝可能在细胞质中液泡的机械和空间分布中发挥作用。