Differential localization of myosin II isoforms in resting and activated osteoclasts.
Osteoclasts resorb bone through a cyclical process of attachment to matrix, polarization, retraction, and migration. Although this process requires major alterations in the organization of actin structures, little is known about roles that myosins play in osteoclast cytoskeletal dynamics. We performed immunolocalization of myosin II using antibodies against heavy chain isoforms IIA and IIB and found that osteoclasts expressed the isoforms in distinct subcellular locations. Myosin IIA was enriched in dynamic cytoskeletal compartments, including the sealing zones of polarized and unpolarized osteoclasts. In contrast, myosin IIB was generally absent from these regions and maintained a comparatively static distribution during different phases of the osteoclast activation cycle. Inhibition of myosin II in osteoclasts by treatment with 2,3-butanedione monoxime caused detachment of unpolarized, but not polarized, cells from the bone matrix. These results suggest that myosin IIA is critical to development of an activated osteoclast phenotype.
Krits
I
I
Renal Division, Washington University School of Medicine, St Louis, MO 63110, USA.
Wysolmerski
R B
RB
Holliday
L S
LS
Lee
B S
BS
D000199:Actins / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D016764:Cell Polarity / Q000187:drug effects; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism; D003931:Diacetyl / Q000494:pharmacology; D007150:Immunohistochemistry; D008264:Macrophages / Q000166:cytology / Q000378:metabolism; D051379:Mice; D024507:Nonmuscle Myosin Type IIA / Q000378:metabolism*; D024501:Nonmuscle Myosin Type IIB / Q000276:immunology / Q000378:metabolism*; D010010:Osteoclasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
D000199:Actins; D024507:Nonmuscle Myosin Type IIA; D024501:Nonmuscle Myosin Type IIB; D003931:Diacetyl