Abstract
Osteoprotegerin (OPG) and the receptor activator of nuclear factor (NF)-kB ligand (RANKL) are key regulators of osteoclastogenesis. The present study had the main aim of showing the localization of OPG and RANKL mRNA and protein in serial sections of the rat femurs and tibiae by immunohistochemistry (IHC) and in situ hybridization (ISH). The main results were: (1) OPG and RANKL mRNA and protein were co-localized in the same cell types, (2) maturative/hypertrophic chondrocytes, osteoblasts, lining cells, periosteal cells and early osteocytes were stained by both IHC and ISH, (3) OPG and RANKL proteins were mainly located in Golgi areas, and the ISH reaction was especially visible in active osteoblasts, (4) immunolabeling was often concentrated into cytoplasmic vacuoles of otherwise negative proliferative chondrocytes; IHC and ISH labeling increased from proliferative to maturative/hypertrophic chondrocytes, (5) the newly laid down bone matrix, cartilage-bone interfaces, cement lines, and trabecular borders showed light OPG and RANKL immunolabeling, (6) about 70% of secondary metaphyseal bone osteocytes showed OPG and RANKL protein expression; most of them were ISH-negative, (7) osteoclasts were mostly unstained by IHC and variably labeled by ISH. The co-expression of OPG and RANKL in the same bone cell types confirms their strictly coupled action in the regulation of bone metabolism.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
DM Anderson E Maraskovsky WL Billingsley WC Dougall ME Tometsko ER Roux MC Teepe RF DuBose D Cosman L Galibert (1997) ArticleTitleA homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function Nature 390 175–179
GJ Atkins DR Haynes SM Geary M Loric TN Crotti DM Findlay (2000) ArticleTitleCoordinated cytokine expression by stromal and hematopoietic cells during human osteoclast formation Bone 26 653–661
N Chikatsu Y Takeuchi S Fukumoto K Yano N Fujita T Tsuruo T Fujita (2002) ArticleTitleClonal endothelial cells produce humoral factors that inhibit osteoclast-like cell formation in vitro J Endocrinol 49 439–447
CR Dhore JP Cleutjens E Lutgens KB Cleutjens PP Geusens PJ Kitslaar JH Tordoir HM Spronk C Vermeer MJ Daemen (2001) ArticleTitleDifferential expression of bone matrix regulatory proteins in human atherosclerotic plaques Arterioscler Thromb Vasc Biol 21 1998–2003
H Enomoto S Shiojiri K Hoshi T Furuichi R Fukuyama CA Yoshida N Kanatani R Nakamura A Mizuno A Zanma K Yano H Yasuda K Higashio K Takada T Komori (2003) ArticleTitleInduction of osteoclast differentiation by Runx2 through receptor activator of nuclear-kB ligand (RANKL) and osteoprotegerin regulation and partial rescue of osteoclastogenesis in Runx2 -/- mice by RANKL transgene J Biol Chem 278 23971–23977
LC Hofbauer S Khosla CR Dunstan DL Lacey WJ Boyle BL Riggs (2000) ArticleTitleThe roles of osteoprotegerin and osteoprotegerin ligand in the paracrine regulation of bone resorption J Bone Min Res 15 2–12
H Hsu DL Lacey CR Dunstan I Solovyev A Colombero E Timms HL Tan G Elliott MJ Kelley I Sarosi L Wang XZ Xia R Elliott L Chiu T Black S Scully C Capparelli S Morony G Shimamoto MB Bass WJ Boyle (1999) ArticleTitleTumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand Proc Natl Acad Sci USA 96 3540–3545
T Ikeda M Utsuyama K Hirokawa (2001) ArticleTitleExpression profiles of receptor activator of nuclear factor kappaB ligand, receptor activator of nuclear factor kappaB, and osteoprotegerin messenger RNA in aged and ovariectomized rat bones J Bone Miner Res 16 1416–1425
V Kartsogiannis H Zhou NJ Horwood RJ Thomas DK Hards JM Quinn P Niforas KW Ng TJ Martin MT Gillespie (1999) ArticleTitleLocalization of RANKL (receptor activator of NF kappa B ligand) mRNA and protein in skeletal and extraskeletal tissues Bone 25 525–534
H Komuro T Olee K Kuhn J Quach DC Brinson A Shikhman J Valbracht L Creighton-Achermann M Lotz (2001) ArticleTitleThe osteoprotegerin/receptor activator of nuclear factor kappaB/receptor activator of nuclear factor kappaB ligand system in cartilage Arthritis Rheum 44 2768–2776
YY Kong H Yoshida I Sarosi HL Tan E Timms C Capparelli S Morony AJ Oliveira-dos-Santos G Van A Itie W Khoo A Wakeham CR Dunstan DL Lacey TW Mak WJ Boyle JM Penninger (1999) ArticleTitleOPGL is a key regulator of osteoclastogenesis, lymphocyte development and lymph-node organogenesis Nature 397 315–323
DL Lacey E Timms HL Tan MJ Kelley CR Dunstan T Burgess R Elliott A Colombero G Elliott S Scully H Hsu J Sullivan N Hawkins E Davy C Capparelli A Eli YX Qian S Kaufman I Sarosi V Shalhoub G Senaldi J Guo J Delaney WJ Boyle (1998) ArticleTitleOsteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation Cell 93 165–176
UM Malyankar M Scatena KL Suchland TJ Yun EA Clark CM Giachelli (2000) ArticleTitleOsteoprotegerin is an αvβ3-induced, NF-kB-dependent survival factor for endothelial cells J Biol Chem 275 20959–20962
H Nakamura T Tsuji A Hirata T Yamamoto (2002) ArticleTitleLocalization of osteoprotegerin (OPG) on bone surfaces and cement lines in rat tibia J Histochem Cytochem 50 945–953
T Okada S Akikusa H Okuno M Kodaka (2003) ArticleTitleBone marrow metastatic myeloma promote osteoclastogenesis through RANKL on endothelial cells Clin Exp Metastasis 20 639–646
JE Onyia RR Miles X Yang DL Halladay J Hale A Glasebrook D McClure G Seno L Churgay S Chandrasekhar TJ Martin (2000) ArticleTitleIn vivo demonstration that human parathyroid hormone 1–38 inhibits the expression of osteoprotegerin in bone with the kinetics of an immediate early gene J Bone Miner Res 15 863–871
T Oshiro Y Shibasaki TJ Martin T Sasaki (2001) ArticleTitleImmunolocalization of vacuolar-type H+-ATPase, cathepsin K, matrix metalloproteinase-9, and receptor activator of NFkappaB ligand in odontoclasts during physiological root resorption of human deciduous teeth Anat Rec 264 305–311
JM Quinn J Elliott MT Gillespie TJ Martin (1998) ArticleTitleA combination of osteoclast differentiation factor and macrophage- colony stimulating factor is sufficient for both human and mouse osteoclast formation in vitro Endocrinology 139 4424–4427
WS Simonet DL Lacey CR Dunstan M Kelley MS Chang R Luthy HQ Nguyen S Wooden L Bennett T Boone G Shimamoto M DeRose R Elliott A Colombero HL Tan G Trail J Sullivan E Davy N Bucay L Renshaw-Gegg TM Hughes D Hill W Pattison P Campbell WJ Boyle (1997a) ArticleTitleOsteoprotegerin: a novel secreted protein involved in the regulation of bone density Cell 89 309–319
WS Simonet DL Lacey CR Dunstan M Kelley MS Chang R Luthy HQ Nguyen S Wooden L Bennett T Boone G Shimamoto M DeRose R Elliott A Colombero HL Tan G Trail J Sullivan E Davy N Bucay L Renshaw-Gegg TM Hughes D Hill W Pattison P Campbell WJ Boyle (1997b) ArticleTitleOsteoprotegerin: a novel secreted protein involved in the regulation of bone density Cell 89 309–319
J Xu JW Tan L Huang XH Gao R Laird D Liu S Wysocki MH Zheng (2000) ArticleTitleCloning, sequencing, and functional characterization of the rat homologue of receptor activator of NF-kappaB ligand J Bone Miner Res 15 2178–2186
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Silvestrini, G., Ballanti, P., Patacchioli, F. et al. Detection of osteoprotegerin (OPG) and its ligand (RANKL) mRNA and protein in femur and tibia of the rat. J Mol Hist 36, 59–67 (2005). https://doi.org/10.1007/s10735-004-3839-1
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/s10735-004-3839-1