Abstract
Osteoclasts are giant bone-resorbing polykaryons that differentiate from mononuclear macrophage/monocyte-lineage hematopoietic precursors. They play critical roles not only in normal bone homeostasis (remodeling) but also in the pathogenesis of bone-destructive disorders such as osteoporosis and rheumatoid arthritis. However, how the activity of mature osteoclasts is regulated in vivo remains unclear. To answer this question, we recently developed an advanced imaging system to visualize living bone tissues with intravital multiphoton microscopy. Using this system, we succeeded in visualization of mature osteoclasts in living bones.
We herein describe the detailed methodology for visualizing bone resorption of mature osteoclasts in living bone marrow and joints using intravital multiphoton microscopy. This approach would be beneficial for studying the cellular dynamics in arthritic inflammation and bone destruction in vivo and would thus be useful for evaluating novel anti-bone-resorptive drugs.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Firestein GS (2003) Evolving concepts of rheumatoid arthritis. Nature 423:356–361
Bromley M, Woolley DE (1984) Histopathology of the rheumatoid lesion. Identification of cell types at sites of cartilage erosion. Arthritis Rheum 27:857–863
Shimizu S, Shiozawa S, Shiozawa K, Imura S, Fujita T (1985) Quantitative histologic studies on the pathogenesis of periarticular osteoporosis in rheumatoid arthritis. Arthritis Rheum 28: 25–31
Teitelbaum SL (2000) Bone resorption by osteoclasts. Science 289:1504–1508
Ishii M et al (2009) Sphingosine-1-phosphate mobilizes osteoclast precursors and regulates bone homeostasis. Nature 458:524–528
Ishii M, Kikuta J, Shimazu Y, Meier-Schellersheim M, Germain RN (2010) Chemorepulsion by blood S1P regulates osteoclast precursor mobilization and bone remodeling in vivo. J Exp Med 207:2793–2798
Cahalan MD, Parker I, Wei SH, Miller MJ (2002) Two-photon tissue imaging: seeing the immune system in a fresh light. Nat Rev Immunol 2:872–880
Germain RN, Miller MJ, Dustin ML, Nussenzweig MC (2006) Dynamic imaging of the immune system: progress, pitfalls and promise. Nat Rev Immunol 6:497–507
Wang BG, Konig K, Halbhuber KJ (2010) Two-photon microscopy of deep intravital tissues and its merits in clinical research. J Microsc 238:1–20
Germain RN et al (2008) Making friends in out-of-the-way places: how cells of the immune system get together and how they conduct their business as revealed by intravital imaging. Immunol Rev 221:163–181
Blair HC, Teitelbaum SL, Ghiselli R, Gluck S (1989) Osteoclastic bone resorption by a polarized vacuolar proton pump. Science 245: 855–857
Nakamura H, Moriyama Y, Futai M, Ozawa H (1994) Immunohistochemical localization of vacuolar H(+)-ATPase in osteoclasts of rat tibiae. Arch Histol Cytol 57:535–539
Toyomura T, Oka T, Yamaguchi C, Wada Y, Futai M (2000) Three subunit a isoforms of mouse vacuolar H(+)-ATPase. Preferential expression of the a3 isoform during osteoclast differentiation. J Biol Chem 275: 8760–8765
Toyomura T et al (2003) From lysosomes to the plasma membrane: localization of vacuolar-type H + -ATPase with the a3 isoform during osteoclast differentiation. J Biol Chem 278: 22023–22030
Sun-Wada GH, Tabata H, Kawamura N, Aoyama M, Wada Y (2009) Direct recruitment of H+-ATPase from lysosomes for phagosomal acidification. J Cell Sci 122:2504–2513
Kikuta J et al (2013) Dynamic visualization of RANKL and Th17-mediated osteoclast function. J Clin Invest 123:866–873
Kong YY et al (1999) Activated T cells regulate bone loss and joint destruction in adjuvant arthritis through osteoprotegerin ligand. Nature 402:304–309
Takayanagi H et al (2000) T-cell-mediated regulation of osteoclastogenesis by signalling cross-talk between RANKL and IFN-gamma. Nature 408:600–605
Sato K et al (2006) Th17 functions as an osteoclastogenic helper T cell subset that links T cell activation and bone destruction. J Exp Med 203:2673–2682
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Kikuta, J., Ishii, M. (2014). Intravital Multiphoton Microscopy for Dissecting Cellular Dynamics in Arthritic Inflammation and Bone Destruction. In: Shiozawa, S. (eds) Arthritis Research. Methods in Molecular Biology, vol 1142. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0404-4_1
Download citation
DOI: https://doi.org/10.1007/978-1-4939-0404-4_1
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-0403-7
Online ISBN: 978-1-4939-0404-4
eBook Packages: Springer Protocols