PMID- 15824857
OWN - NLM
STAT- MEDLINE
DCOM- 20050819
LR  - 20091119
IS  - 0884-0431 (Print)
IS  - 0884-0431 (Linking)
VI  - 20
IP  - 5
DP  - 2005 May
TI  - TRAF2 is essential for TNF-alpha-induced osteoclastogenesis.
PG  - 840-7
AB  - TRAF2-deficient mice show embryonic lethality, and we developed a new in vitro 
      differentiation system to show the function of TRAF2 in osteoclastogenesis, in 
      which osteoclast progenitors are derived from the fetal liver of TRAF2-deficient 
      mice. Using this system, we showed that TRAF2 is required for TNF-alpha-induced 
      osteoclastogenesis. INTRODUCTION: TNF receptor-associated factor 2 (TRAF2) is a 
      signal transducer for RANK and for two TNF receptor isotypes, TNFR1 and TNFR2. 
      Because TRAF2-deficient mice show embryonic lethality, it has remained unclear 
      whether TRAF2 is crucial in RANKL- or TNF-alpha-induced osteoclastogenesis. 
      MATERIALS AND METHODS: Osteoclast progenitors derived from fetal liver were 
      cultured in the presence of monocyte macrophage colony-stimulating factor 
      (M-CSF), and flow cytometry for characterization of surface markers on these 
      cells was performed. To examine the involvement of TRAF2 in osteoclast 
      differentiation, we cultured osteoclast progenitors from TRAF2-deficient and 
      wildtype mice with soluble RANKL or TNF-alpha in the presence of M-CSF, and 
      counted the number of TRACP(+) multinucleate cells formed. c-jun N-terminal 
      kinase (JNK) and NF-kappaB activation in osteoclast progenitors was examined by 
      Western blot analysis and electrophoretic mobility shift assay, respectively. 
      Nuclear factor of activated T cells (NFATc1) expression and activation were 
      analyzed by RT-PCR and immunofluorescence staining, respectively. To examine 
      whether TRAF2 overexpression induced osteoclastogenesis, TRAF2 was overexpressed 
      in osteoclast progenitors form wildtype bone marrow by retrovirus infection. 
      RESULTS AND CONCLUSIONS: Osteoclast progenitors from normal fetal liver, which 
      were cultured with M-CSF, expressed surface molecules c-fms, Mac-1, and RANK, and 
      could differentiate into TRACP(+) multinucleate cells in the presence of soluble 
      RANKL or TNF-alpha. RANKL-induced osteoclastogenesis gave a reduction of 20% in 
      the progenitors from TRAF2-deficient mice compared with that of the cells from 
      littermate wildtype mice, whereas TNF-alpha-induced osteoclastogenesis was 
      severely impaired in the cells from the TRAF2-deficient mice. Only a few TRACP(+) 
      multinucleate cells were formed, and TNF-alpha-mediated activation of JNK, 
      NF-kappaB, and NFATc1 was defective. TRAF2 overexpression induced differentiation 
      of osteoclast progenitors from wildtype mice into TRACP(+) multinucleate cells. 
      These results suggest that TRAF2 plays an important role in TNF-alpha-induced 
      osteoclastogenesis.
FAU - Kanazawa, Kiyoshi
AU  - Kanazawa K
AD  - Department of Life Science, Tokyo Institute of Technology, Yokohama, Japan.
FAU - Kudo, Akira
AU  - Kudo A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20041220
PL  - England
TA  - J Bone Miner Res
JT  - Journal of bone and mineral research : the official journal of the American 
      Society for Bone and Mineral Research
JID - 8610640
RN  - 0 (Carrier Proteins)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (NF-kappa B)
RN  - 0 (NFATC Transcription Factors)
RN  - 0 (Nfatc1 protein, mouse)
RN  - 0 (Nuclear Proteins)
RN  - 0 (RANK Ligand)
RN  - 0 (Receptor Activator of Nuclear Factor-kappa B)
RN  - 0 (TNF Receptor-Associated Factor 2)
RN  - 0 (Tnfrsf11a protein, mouse)
RN  - 0 (Tnfsf11 protein, mouse)
RN  - 0 (Transcription Factors)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 81627-83-0 (Macrophage Colony-Stimulating Factor)
RN  - EC 2.7.11.24 (JNK Mitogen-Activated Protein Kinases)
RN  - EC 2.7.12.2 (MAP Kinase Kinase 4)
RN  - EC 2.7.12.2 (Mitogen-Activated Protein Kinase Kinases)
SB  - IM
MH  - Animals
MH  - Blotting, Western
MH  - Bone Marrow Cells/cytology
MH  - Bone Resorption
MH  - Carrier Proteins/metabolism
MH  - Cell Differentiation
MH  - Cell Nucleus/metabolism
MH  - Cells, Cultured
MH  - DNA-Binding Proteins/metabolism
MH  - Flow Cytometry
MH  - Genotype
MH  - JNK Mitogen-Activated Protein Kinases/metabolism
MH  - Liver/cytology/embryology
MH  - MAP Kinase Kinase 4
MH  - Macrophage Colony-Stimulating Factor/metabolism
MH  - Male
MH  - Membrane Glycoproteins/metabolism
MH  - Mice
MH  - Mice, Transgenic
MH  - Microscopy, Fluorescence
MH  - Mitogen-Activated Protein Kinase Kinases/metabolism
MH  - NF-kappa B/metabolism
MH  - NFATC Transcription Factors
MH  - Nuclear Proteins/metabolism
MH  - Osteoclasts/*cytology/metabolism
MH  - Plasmids/metabolism
MH  - RANK Ligand
MH  - Receptor Activator of Nuclear Factor-kappa B
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Signal Transduction
MH  - Stem Cells/cytology
MH  - TNF Receptor-Associated Factor 2/*physiology
MH  - Time Factors
MH  - Transcription Factors/metabolism
MH  - Transgenes
MH  - Tumor Necrosis Factor-alpha/*metabolism
EDAT- 2005/04/13 09:00
MHDA- 2005/08/20 09:00
CRDT- 2005/04/13 09:00
PHST- 2004/08/23 00:00 [received]
PHST- 2004/11/10 00:00 [revised]
PHST- 2004/12/14 00:00 [accepted]
PHST- 2005/04/13 09:00 [pubmed]
PHST- 2005/08/20 09:00 [medline]
PHST- 2005/04/13 09:00 [entrez]
AID - 10.1359/JBMR.041225 [doi]
PST - ppublish
SO  - J Bone Miner Res. 2005 May;20(5):840-7. doi: 10.1359/JBMR.041225. Epub 2004 Dec 
      20.