A new method to isolate large numbers of rabbit osteoclasts and osteoclast-like cells: application to the characterization of serum response element binding proteins during osteoclast differentiation.
We have developed a new method that allows the purification of large numbers of both authentic osteoclasts (OCs) and in vitro differentiated osteoclast-like cells (OCLs) from rabbits. We characterized the OCLs in terms of the expression of different phenotypic markers of OC differentiation and their ability to resorb bone. The method provides a system for performing biochemical and molecular studies of OC differentiation and function in a single species. We used this system to characterize the effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on the expression of proteins that bind to the serum response element (SRE) of the c-fos promoter. We found that OCLs and OCs displayed similar SRE-binding activities, including the serum response factor (SRF). This pattern is established in a time-dependent and cell-specific manner in response to long-term treatment of rabbit bone marrow by 1,25(OH)2D3. Thus, 1,25(OH)2D3 can modulate SRF and/or SRF-related protein. This finding may contribute to understanding the role of c-Fos in the regulation of OC differentiation.
David
J P
JP
Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520-8044, USA.
Neff
L
L
Chen
Y
Y
Rincon
M
M
Horne
W C
WC
Baron
R
R
D000818:Animals; D000831:Animals, Newborn; D001798:Blood Proteins / Q000378:metabolism; D001862:Bone Resorption; D002116:Calcitonin / Q000494:pharmacology; D002117:Calcitriol / Q000494:pharmacology*; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods*; D018920:Coculture Techniques; D007150:Immunohistochemistry; D010010:Osteoclasts / Q000166:cytology / Q000502:physiology*; D016760:Proto-Oncogene Proteins c-fos / Q000502:physiology*; D011817:Rabbits
D001798:Blood Proteins; D016760:Proto-Oncogene Proteins c-fos; D002116:Calcitonin; D002117:Calcitriol