Immunohistochemical localization of vacuolar H(+)-ATPase in osteoclasts of rat tibiae.
The localization of vacuolar H(+)-atpASE (V-ATPase) in osteoclasts of rat tibiae was examined immunohistochemically using antibodies to 57 kD and 72 kD subunits of V-ATPase, with confocal laser scanning microscopy and transmission electron microscopy. The two antibodies showed similar immunoreactivities in osteoclasts. We distinguished three patterns of immunoreactivity in osteoclasts: The first pattern was that immunoreactivity was diffusely detected in cytoplasm of osteoclasts; The second one was that osteoclasts showed intense immunoreactivity in their regions contacting with bone surface; and the last was that little immunoreactivity was seen in osteoclasts. Post-embedding methods with protein A-gold complex revealed that the subunits of V-ATPase were localized not only on the membranes of the ruffled borders of osteoclasts, but also around the Golgi apparatus and accumulated tubular lysosomes. These findings suggest that: 1) immunoreactive patterns for V-ATPase in osteoclasts reflect the polarity and activity of osteoclasts; and 2) V-ATPase on ruffled borders may be transported through tubular lysosomes.
Nakamura
H
H
Department of Oral Anatomy, Niigata University School of Dentistry, Japan.
Moriyama
Y
Y
Futai
M
M
Ozawa
H
H
D000818:Animals; D007150:Immunohistochemistry; D008297:Male; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D010010:Osteoclasts / Q000737:chemistry* / Q000201:enzymology / Q000648:ultrastructure; D006180:Proton-Translocating ATPases / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D013977:Tibia / Q000166:cytology*; D014617:Vacuoles / Q000737:chemistry* / Q000201:enzymology*
D006180:Proton-Translocating ATPases