Ultrastructural analysis of apatite-degrading capability of extended invasive podosomes in resorbing osteoclasts.
Osteoclasts in culture are non-transformed cell types that spontaneously develop specific cell-adhesion devices such as podosomes. An individual podosome is a complex network of filamentous actin (F-actin) unit structure that collectively, with other proteins, self-organizes as the sealing zone. Major matrix degradation on apatite seems to proceed under the ruffled-border domain, which is an enclosed extracellular compartment tightly sealed off by this sealing zone. Presently we found that usually the top of finger-like projections of the ruffled border reached toward the plane of the apatite surface, where a shallow degradation of apatite took place. Simultaneously, we obtained several pieces of structural evidence indicating that a specific protrusion referred to as an invasive podosome (invadopodium), which was continuous with podosomes derived from the sealing zone, invaded deeply into apatite matrix and degraded it. The F-actin architecture of the invasive podosome - an active extracellular matrix-degrading, actin-rich cell protrusion - could be distinguished from that of other punctate F-actin structures including the individual podosome, sealing zone, and ruffled border projection. Invasive podosomes contained 2 different F-actin populations, i.e., an interconnected meshwork and a parallel array of bundles. The morphological variability of these protrusions was apparent, having a single cylindrical to lamella-shaped cytoskeletal organization. Our present observations strongly suggest that the degradation of apatite substrate-resorbing osteoclasts appears to have been preceded by the combined appearance of ruffled border and invasive podosomes, and also occurred simultaneously with cell migration during an alternating cycle of resorption and migration.
Department of Oral Anatomy and Neurobiology, Osaka University Graduate School of Dentistry, Yamadaoka 1-8, Suita, Osaka 565-0871, Japan. Electronic address: takisaka@dent.osaka-u.ac.jp.
Toshitaka
T
Akisaka
Department of Oral Anatomy and Neurobiology, Osaka University Graduate School of Dentistry, Yamadaoka 1-8, Suita, Osaka 565-0871, Japan. Electronic address: yoshida@dent.osaka-u.ac.jp.
Astushi
A
Yoshida
D008841:Actin Cytoskeleton / Q000648:ultrastructure; D000818:Animals; D001031:Apatites / Q000378:metabolism*; D001862:Bone Resorption; D002448:Cell Adhesion; D021961:Cell Membrane Structures / Q000648:ultrastructure; D002465:Cell Movement; D002478:Cells, Cultured; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D010010:Osteoclasts / Q000378:metabolism / Q000648:ultrastructure*; D000069261:Podosomes / Q000378:metabolism* / Q000648:ultrastructure*; D011817:Rabbits; D016628:Replica Techniques
D001031:Apatites
Anaglyph stereo; Invasive podosome; Replica microscopy; Resorbing osteoclast; Ultrastructure