title: Normal human osteoclasts formed from peripheral blood monocytes express PTH type 1 receptors and are stimulated by PTH in the absence of osteoblasts.
abstract: The prevailing view for many years has been that osteoclasts do not express parathyroid hormone (PTH) receptors and that PTH's effects on osteoclasts are mediated indirectly via osteoblasts. However, several recent reports suggest that osteoclasts express PTH receptors. In this study, we tested the hypothesis that human osteoclasts formed in vitro express functional PTH type 1 receptors (PTH1R). Peripheral blood monocytes (PBMC) were cultured on bone slices or plastic culture dishes with human recombinant RANK ligand (RANKL) and recombinant human macrophage colony-stimulating factor (M-CSF) for 16-21 days. This resulted in a mixed population of mono- and multi-nucleated cells, all of which stained positively for the human calcitonin receptor. The cells actively resorbed bone, as assessed by release of C-terminal telopeptide of type I collagen and the formation of abundant resorption pits. We obtained evidence for the presence of PTH1R in these cells by four independent techniques. First, using immunocytochemistry, positive staining for PTH1R was observed in both mono- and multi-nucleated cells intimately associated with resorption cavities. Second, PTH1R protein expression was demonstrated by Western blot analysis. Third, the cells expressed PTH1R mRNA at 21 days and treatment with 10(-7) M hPTH (1-34) reduced PTH1R mRNA expression by 35%. Finally, bone resorption was reproducibly increased by two to threefold when PTH (1-34) was added to the cultures. These findings provide strong support for a direct stimulatory action of PTH on human osteoclasts mediated by PTH1R. This suggests a dual regulatory mechanism, whereby PTH acts both directly on osteoclasts and also, indirectly, via osteoblasts.
authors.affiliation: Regional Bone Center, Helen Hayes Hospital, New York State Department of Health, West Haverstraw, New York 10993, USA. dempster@helenhayeshosp.org
authors.forename: David W
authors.initials: DW
authors.lastname: Dempster
authors.forename: Christine E
authors.initials: CE
authors.lastname: Hughes-Begos
authors.forename: Katarina
authors.initials: K
authors.lastname: Plavetic-Chee
authors.forename: Andrea
authors.initials: A
authors.lastname: Brandao-Burch
authors.forename: Felicia
authors.initials: F
authors.lastname: Cosman
authors.forename: Jeri
authors.initials: J
authors.lastname: Nieves
authors.forename: Simon
authors.initials: S
authors.lastname: Neubort
authors.forename: Shi Shou
authors.initials: SS
authors.lastname: Lu
authors.forename: Akiko
authors.initials: A
authors.lastname: Iida-Klein
authors.forename: Tim
authors.initials: T
authors.lastname: Arnett
authors.forename: Robert
authors.initials: R
authors.lastname: Lindsay
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000502:physiology; D009000:Monocytes / Q000166:cytology / Q000502:physiology*; D010006:Osteoblasts / Q000378:metabolism; D010010:Osteoclasts / Q000166:cytology / Q000378:metabolism*; D010281:Parathyroid Hormone / Q000494:pharmacology*; D044168:Receptor, Parathyroid Hormone, Type 1 / Q000096:biosynthesis*
chemical_list: D010281:Parathyroid Hormone; D044168:Receptor, Parathyroid Hormone, Type 1