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Blockade of the Pore-Forming P2X7 Receptor Inhibits Formation of Multinucleated Human Osteoclasts In Vitro

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Abstract

Osteoclasts are large, multinucleated, terminally differentiated cells formed by the fusion of mononuclear hemopoietic precursors. Their function is the resorption of bone, which is an essential part of the growth, modeling and remodeling of the skeleton. Though some osteoclast differentiation factors have recently been identified, the molecular basis for the fusion process that leads to multinucleation is poorly understood. The ATP-gated P2X7 receptor is a plasma membrane receptor belonging to the family of P2X purinergic receptors. It is known to be expressed by cells of hemopoietic origin where its activation leads to multiple downstream events including cytokine release, cell permeabilization and apoptosis. More recently this receptor has been implicated in the generation of multinucleated giant cells and polykaryons. Here we show that human osteoclasts express P2X7 receptors in vitro and in vivo, and that these receptors are functional in vitro, as assessed by pore-formation studies. More importantly, blockade of the P2X7 receptor with the antagonist oxidized ATP or a blocking monoclonal antibody significantly inhibits the fusion of osteoclast precursors to form multinucleated osteoclasts. Taken in combination with previous results from our laboratory demonstrating P2X7 receptor-mediated apoptosis and inhibition of bone resorption in vitro, these data suggest an important role for the P2X7 receptor in the regulation of the osteoclast population. The P2X7 receptor provides a significant new target for modulating osteoclast function in diseases characterized by increased osteoclast number and excessive bone turnover.

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Acknowledgements

We are grateful to Prof. W.D. Fraser for supplying samples from patients under his care; J. Rice for help with the ethidium bromide uptake images; Drs. Odgren, Marks and Kramer-Haemmerle for helpful discussions, and Dr. Ivings for assistance with the manuscript. This work was supported by The University of Liverpool (A.G.), and the Arthritis Research Campaign (K.A.B.).

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Authors and Affiliations

  1. Human Bone Cell Research Group, Department Human Anatomy and Cell Biology, The University of Liverpool, Liverpool, L69 3GE, UK

    A. Gartland, K. A. Buckley, W. B. Bowler & J. A. Gallagher

  2. Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Avenue N., Worcester, Massachusetts, 01655, USA

    A. Gartland

  3. Strakan Pharmaceuticals, Buckholm Mill Brae, Buckholm Mill, Galashiels, TD1 2HB, UK

    W. B. Bowler

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  1. A. Gartland
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  2. K. A. Buckley
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  4. J. A. Gallagher
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Correspondence to A. Gartland.

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Gartland, A., Buckley, K., Bowler, W. et al. Blockade of the Pore-Forming P2X7 Receptor Inhibits Formation of Multinucleated Human Osteoclasts In Vitro . Calcif Tissue Int 73, 361–369 (2003). https://doi.org/10.1007/s00223-002-2098-y

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