NF-kappaB p50 and p52 expression is not required for RANK-expressing osteoclast progenitor formation but is essential for RANK- and cytokine-mediated osteoclastogenesis

J Bone Miner Res. 2002 Jul;17(7):1200-10. doi: 10.1359/jbmr.2002.17.7.1200.

Abstract

Expression of RANKL by stromal cells and of RANK and both NF-kappaB p50 and p52 by osteoclast precursors is essential for osteoclast formation. To examine further the role of RANKL, RANK, and NF-KB signaling in this process, we used NF-kappaB p50-/- ;p52-/- double knockout (dKO) and wild-type (WT) mice. Osteoclasts formed in cocultures of WT osteoblasts with splenocytes from WT mice but not from dKO mice, a finding unchanged by addition of RANKL and macrophage colony-stimulating factor (M-CSF). NF-kappaB dKO splenocytes formed more colony-forming unit granulocyte macrophage (CFU-GM) colonies than WT cells, but no osteoclasts were formed from dKO CFU-GM colonies. RANKL increased the number of CFU-GM colonies twofold in WT cultures but not in dKO cultures. Fluorescence-activated cell sorting (FACS) analysis of splenocytes from NF-kappaB dKO mice revealed a two-to threefold increase in the percentage of CD11b (Mac-1) and RANK double-positive cells compared with WT controls. Treatment of NF-kappaB dKO splenocytes with interleukin (IL)-1, TNF-alpha, M-CSF, GM-CSF, and IL-6 plus soluble IL-6 receptor did not rescue the osteoclast defect. No increase in apoptosis was observed in cells of the osteoclast lineage in NF-kappaB dKO or p50-/-;p52+/- (3/4KO) mice. Thus, NF-kappaB p50 and p52 expression is not required for formation of RANK-expressing osteoclast progenitors but is essential for RANK-expressing osteoclast precursors to differentiate into TRAP+ osteoclasts in response to RANKL and other osteoclastogenic cytokines.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antioxidants / pharmacology
  • Apoptosis
  • Bone Marrow Cells / cytology
  • Carrier Proteins / pharmacology
  • Cell Differentiation
  • Cells, Cultured
  • Glycoproteins / biosynthesis*
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Interleukin-1 / pharmacology
  • Interleukin-6 / pharmacology
  • Leupeptins / pharmacology
  • Macrophage Colony-Stimulating Factor / pharmacology
  • Membrane Glycoproteins / pharmacology
  • Mice
  • Mice, Knockout
  • NF-kappa B / antagonists & inhibitors
  • NF-kappa B / genetics
  • NF-kappa B / metabolism*
  • NF-kappa B p50 Subunit
  • Osteoclasts / cytology*
  • Osteoclasts / drug effects
  • Osteoclasts / metabolism
  • Osteoprotegerin
  • Proline / analogs & derivatives*
  • Proline / pharmacology
  • RANK Ligand
  • Receptor Activator of Nuclear Factor-kappa B
  • Receptors, Cytoplasmic and Nuclear / biosynthesis*
  • Receptors, Interleukin-6
  • Receptors, Tumor Necrosis Factor / biosynthesis*
  • Signal Transduction*
  • Spleen / cytology
  • Spleen / metabolism
  • Stem Cells / cytology*
  • Stem Cells / drug effects
  • Stem Cells / metabolism
  • Stromal Cells / cytology
  • Stromal Cells / drug effects
  • Thiocarbamates / pharmacology
  • Tosylphenylalanyl Chloromethyl Ketone / pharmacology
  • Tumor Necrosis Factor-alpha / pharmacology

Substances

  • Antioxidants
  • Carrier Proteins
  • Glycoproteins
  • Interleukin-1
  • Interleukin-6
  • Leupeptins
  • Membrane Glycoproteins
  • NF-kappa B
  • NF-kappa B p50 Subunit
  • Osteoprotegerin
  • RANK Ligand
  • Receptor Activator of Nuclear Factor-kappa B
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Interleukin-6
  • Receptors, Tumor Necrosis Factor
  • Thiocarbamates
  • Tnfrsf11a protein, mouse
  • Tnfrsf11b protein, mouse
  • Tnfsf11 protein, mouse
  • Tumor Necrosis Factor-alpha
  • prolinedithiocarbamate
  • Tosylphenylalanyl Chloromethyl Ketone
  • Macrophage Colony-Stimulating Factor
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Proline
  • benzyloxycarbonylleucyl-leucyl-leucine aldehyde