Abstract
Macrophages and osteoclasts are thought to derive from CD68 lineage marker-positive common myeloid precursors. We used the CD68 promoter to drive an inducible receptor activator of NF-κB (iRANK) construct that selectively activates RANK signaling in myeloid cells in vivo. The cytoplasmic portion of RANK was fused to a mutant FK506 binding domain, which selectively binds the chemical inducer of dimerization AP20187 and initiates signaling. iRANK mRNA was expressed in macrophages isolated from peritoneal cavity, spleen-, and bone marrow-derived myeloid cells. Unexpectedly, AP20187 did not induce osteoclast formation in spleen- and bone marrow-derived myeloid cells. However, AP20187-dependent RANK signaling induced ERK1/2 phosphorylation and mRNA expression of MMP9 and CathepsinK in peritoneal macrophages. Importantly, CD68 was not expressed until day 3 and day 5 in bone marrow and spleen myeloid cells, respectively. Contrary to dogma, osteoclast precursors do not express the lineage marker CD68.
Keywords:
RANK; CD68; osteoclast precursors.
© 2017 Federation of European Biochemical Societies.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, N.I.H., Extramural
MeSH terms
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Animals
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Antigens, CD / genetics*
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Antigens, CD / metabolism
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Antigens, Differentiation, Myelomonocytic / genetics*
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Antigens, Differentiation, Myelomonocytic / metabolism
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Bone Marrow Cells / cytology
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Bone Marrow Cells / drug effects
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Bone Marrow Cells / metabolism
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Cathepsin K / genetics
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Cathepsin K / metabolism
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Cell Differentiation / drug effects
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Gene Expression Regulation
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Macrophages / cytology
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Macrophages / drug effects
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Macrophages / metabolism*
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Matrix Metalloproteinase 9 / genetics
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Matrix Metalloproteinase 9 / metabolism
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Mice
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Mice, Transgenic
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Mitogen-Activated Protein Kinase 1 / genetics
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Mitogen-Activated Protein Kinase 1 / metabolism
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Mitogen-Activated Protein Kinase 3 / genetics
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Mitogen-Activated Protein Kinase 3 / metabolism
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Myeloid Cells / cytology
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Myeloid Cells / drug effects
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Myeloid Cells / metabolism*
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Osteoclasts / cytology
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Osteoclasts / drug effects
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Osteoclasts / metabolism*
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Peritoneal Cavity / cytology
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Promoter Regions, Genetic
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Receptor Activator of Nuclear Factor-kappa B / genetics*
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Receptor Activator of Nuclear Factor-kappa B / metabolism
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Signal Transduction
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Spleen / cytology
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Spleen / drug effects
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Spleen / metabolism
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Tacrolimus / analogs & derivatives
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Tacrolimus / pharmacology
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Tacrolimus Binding Proteins / genetics
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Tacrolimus Binding Proteins / metabolism
Substances
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AP20187
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Antigens, CD
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Antigens, Differentiation, Myelomonocytic
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CD68 protein, mouse
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Receptor Activator of Nuclear Factor-kappa B
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Recombinant Fusion Proteins
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Tnfrsf11a protein, mouse
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Mapk1 protein, mouse
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Mitogen-Activated Protein Kinase 1
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Mitogen-Activated Protein Kinase 3
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Cathepsin K
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Ctsk protein, mouse
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Matrix Metalloproteinase 9
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Mmp9 protein, mouse
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Tacrolimus Binding Proteins
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Tacrolimus