Osteoclasts express the B2 isoform of vacuolar H(+)-ATPase intracellularly and on their plasma membranes.
Osteoclasts express high levels of vacuolar H(+)-ATPase (V-ATPase) in their ruffled membranes, driving the secretion of H+ required for normal bone resorption. Previous reports have suggested that the B subunit of the osteoclast V-ATPase differs from those expressed in kidney and other tissues. In this study, B subunit isoform-specific antibodies and cDNA probes were used to examine which B subunit isoform is expressed in osteoclasts and osteoclast-like cells. Immunoblotting and RNA hybridization analysis were used to demonstrate that cells from an osteoclast-rich mouse bone marrow culture model express the B2 but not the B1 subunit isoform. Immunocytochemical staining of murine osteoclasts generated in vitro and of native rat osteoclasts in bone sections showed that the B2 but not the B1 isoform was expressed at high levels and was polarized to the ruffled membrane. Human marrow cultures and monocyte-derived macrophages, used as models for osteoclasts, also expressed the B2 but not the B1 subunit isoform. These results indicate that V-ATPases containing the B2 subunit isoform mediate osteoclast bone resorption.
Lee
B S
BS
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Holliday
L S
LS
Ojikutu
B
B
Krits
I
I
Gluck
S L
SL
D000818:Animals; D001483:Base Sequence; D001853:Bone Marrow / Q000201:enzymology; D001854:Bone Marrow Cells; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007425:Intracellular Membranes / Q000201:enzymology; D007527:Isoenzymes / Q000378:metabolism*; D008264:Macrophages / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D009000:Monocytes / Q000201:enzymology; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes / Q000235:genetics; D010010:Osteoclasts / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D012313:RNA / Q000235:genetics; D014617:Vacuoles / Q000201:enzymology*
D007527:Isoenzymes; D015345:Oligonucleotide Probes; D012313:RNA; D006180:Proton-Translocating ATPases