Yasuda, Proc Natl Acad Sci U S A 1998 - Osteoclast Differentiation Factor Is a Ligand for Osteoprotegerin/Osteoclastogenesis-Inhibitory Factor and Is Identical to TRANCE/RANKL
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Osteoclast Differentiation Factor Is a Ligand for Osteoprotegerin/Osteoclastogenesis-Inhibitory Factor and Is Identical to TRANCE/RANKL
Yasuda, Suda, et al.·Proc Natl Acad Sci U S A·1998·4,239 citations·Basic science
Yasuda and colleagues set out to clone the cell‑surface partner of OPG/OCIF from mouse stromal cells and to test whether it drives osteoclast formation.
In vitro study: expression cloning from a mouse stromal‑cell cDNA library, then osteoclast‑formation assays in mouse spleen cell cultures.
Methods
Model
Mouse ST2 stromal cells
Mouse spleen cells as osteoclast progenitors
Primary calvarial osteoblasts from newborn ddY mice
COS‑7 cells for expression cloning
Intervention
COS‑7 cells transfected with ODF expression vector
Soluble ODF (sODF) with M‑CSF added to spleen or C7 cells
OPG/OCIF added to block ODF‑induced osteoclastogenesis
1,25(OH)₂D₃, PTH/PGE₂ or IL‑11 applied to stromal cells and osteoblasts
Assays
TRAP‑positive multinucleated cell formation
Resorption pits on dentine slices
Coimmunoprecipitation with OPG/OCIF
ODF mRNA expression
Time points
6‑day spleen cell cultures with sODF
Key Findings
The cloned protein, named ODF, is a membrane‑bound member of the TNF ligand family that drove osteoclast formation from progenitors.
Soluble ODF plus M‑CSF, without osteoblasts or stromal cells, produced giant TRAP‑positive cells up to 500 µm across from spleen cells, and these cells resorbed dentine.
Adding OPG/OCIF abolished the osteoclast formation that ODF induced.
ODF mRNA was absent in untreated ST2 stromal cells and induced by 1,25(OH)₂D₃ with dexamethasone, and in primary osteoblasts it was up‑regulated by 1,25(OH)₂D₃, IL‑11, PGE₂ or PTH.
ODF is a 316‑amino‑acid type II transmembrane protein with a predicted Mr of 36K, seen as a 40‑kDa protein on coimmunoprecipitation.
Its amino‑acid sequence matched TRANCE/RANKL, a ligand that promotes T‑cell growth and the function of dendritic cells.
Board pearl
Bone‑resorbing factors induce osteoblasts and stromal cells to express membrane‑bound ODF (RANKL), which signals progenitors to become osteoclasts, and secreted OPG binds it to stop that differentiation.
Authors' conclusion
We conclude that the membrane-bound protein is osteoclast differentiation factor (ODF), a long-sought ligand mediating an essential signal to osteoclast progenitors for their differentiation into osteoclasts.
Yasuda and colleagues set out to clone the cell‑surface partner of OPG/OCIF from mouse stromal cells and to test whether it drives osteoclast formation.
In vitro study: expression cloning from a mouse stromal‑cell cDNA library, then osteoclast‑formation assays in mouse spleen cell cultures.
Methods
Model
Mouse ST2 stromal cells
Mouse spleen cells as osteoclast progenitors
Primary calvarial osteoblasts from newborn ddY mice
COS‑7 cells for expression cloning
Intervention
COS‑7 cells transfected with ODF expression vector
Soluble ODF (sODF) with M‑CSF added to spleen or C7 cells
OPG/OCIF added to block ODF‑induced osteoclastogenesis
1,25(OH)₂D₃, PTH/PGE₂ or IL‑11 applied to stromal cells and osteoblasts
Assays
TRAP‑positive multinucleated cell formation
Resorption pits on dentine slices
Coimmunoprecipitation with OPG/OCIF
ODF mRNA expression
Time points
6‑day spleen cell cultures with sODF
Key Findings
The cloned protein, named ODF, is a membrane‑bound member of the TNF ligand family that drove osteoclast formation from progenitors.
Soluble ODF plus M‑CSF, without osteoblasts or stromal cells, produced giant TRAP‑positive cells up to 500 µm across from spleen cells, and these cells resorbed dentine.
Adding OPG/OCIF abolished the osteoclast formation that ODF induced.
ODF mRNA was absent in untreated ST2 stromal cells and induced by 1,25(OH)₂D₃ with dexamethasone, and in primary osteoblasts it was up‑regulated by 1,25(OH)₂D₃, IL‑11, PGE₂ or PTH.
ODF is a 316‑amino‑acid type II transmembrane protein with a predicted Mr of 36K, seen as a 40‑kDa protein on coimmunoprecipitation.
Its amino‑acid sequence matched TRANCE/RANKL, a ligand that promotes T‑cell growth and the function of dendritic cells.
Board pearl
Bone‑resorbing factors induce osteoblasts and stromal cells to express membrane‑bound ODF (RANKL), which signals progenitors to become osteoclasts, and secreted OPG binds it to stop that differentiation.
Authors' conclusion
We conclude that the membrane-bound protein is osteoclast differentiation factor (ODF), a long-sought ligand mediating an essential signal to osteoclast progenitors for their differentiation into osteoclasts.