The intermediate filament cytoskeleton of macrophages

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.