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Updated: Jul 28, 2026
09:37
A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
Published on: March 15, 2018
Expression of functional RANK on mature rat and human osteoclasts.
D E Myers1, F M Collier, C Minkin
1Department of Medicine, The University of Melbourne, Barwon Health, The Geelong Hospital, Geelong, Australia.
FEBS Letters
|December 22, 1999
Summary
The receptor activator of nuclear factor kappa-B (RANK) is highly expressed in mature osteoclasts. Its stimulation by RANK ligand (RANKL) activates NF-kappaB and calcium signaling pathways.
Area of Science:
- Cell Biology
- Molecular Biology
- Bone Biology
Background:
- The roles of RANK/RANKL in osteoclastogenesis are established.
- Their regulatory roles in mature osteoclasts are uncertain.
Purpose of the Study:
- To investigate the expression and function of RANK/RANKL in mature osteoclasts.
- To determine the signaling pathways activated by RANKL in mature osteoclasts.
Main Methods:
- Microisolation of pure rat and human osteoclast populations.
- RT-PCR analysis for gene expression (RANK, calcitonin receptor, OPG, ALP, RANKL).
- Treatment of osteoclasts with soluble RANKL and analysis of NF-kappaB and calcium levels.
Main Results:
- RANK and calcitonin receptor mRNA were detected in all samples.
- OPG and ALP mRNA were absent.
- RANKL mRNA was detected in a subset of rat and human samples.
- Soluble RANKL treatment induced NF-kappaB nuclear translocation and elevated cytosolic and nuclear calcium levels.
Conclusions:
- RANK is highly expressed in mature osteoclasts.
- RANKL stimulation activates NF-kappaB and calcium signaling in mature osteoclasts.
- These findings elucidate novel roles for RANK/RANKL in mature osteoclast regulation.
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