title: Effects of synthetic peptido-leukotrienes on bone resorption in vitro.
abstract: Peptido-leukotrienes are short-lived organic molecules known to have potent biological effects as mediators of inflammation, hypersensitivity and respiratory disorders. However, little is known concerning their effects on bone cells. We have shown previously that stromal cells isolated from a human giant cell tumor secrete 5-HETE (5-hydroxyeicosatetraenoic acid) and the peptido-leukotrienes, also known as the cysteinyl leukotrienes LTC4, LTD4, and LTE4. These eicosanoids were shown to stimulate the multinucleated giant cells obtained from these tumors to form resorption lacunae on sperm whale dentine. Here, we show that the peptido-leukotrienes also stimulate isolated avian osteoclast-like cells to form resorption lacunae and to increase their content of tartrate-resistant acid phosphatase. LTD4 increased 45Ca release from murine calvarial bone organ cultures, but not from fetal rat long bone cultures. Isolated avian osteoclast-like cells were chosen to perform receptor binding studies, as this population is the most homogeneous source of osteoclasts available. After the precursors had fused to form multinucleated cells, receptor binding assays were performed. Scatchard analysis of saturation binding data showed a single class of binding sites, with a dissociation constant (Kd) of 0.53 nM and a receptor density of 5,200 receptors per cell. Competition binding studies showed receptor specificity using a specific LTD4 receptor antagonist ZM 198,615. These data show that the peptido-leukotrienes activate highly enriched populations of isolated avian osteoclast-like cells, and also that specific LTD4 receptors are present in this cell population.
authors.affiliation: University of Texas Health Science Center at San Antonio, Department of Medicine, USA.
authors.forename: C
authors.initials: C
authors.lastname: Garcia
authors.forename: M
authors.initials: M
authors.lastname: Qiao
authors.forename: D
authors.initials: D
authors.lastname: Chen
authors.forename: M
authors.initials: M
authors.lastname: Kirchen
authors.forename: W
authors.initials: W
authors.lastname: Gallwitz
authors.forename: G R
authors.initials: GR
authors.lastname: Mundy
authors.forename: L F
authors.initials: LF
authors.lastname: Bonewald
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D001859:Bone Neoplasms / Q000473:pathology; D001862:Bone Resorption / Q000139:chemically induced*; D001842:Bone and Bones / Q000187:drug effects / Q000503:physiopathology; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D002645:Chickens; D005260:Female; D018212:Giant Cell Tumor of Bone / Q000473:pathology; D006801:Humans; D007191:Indazoles / Q000378:metabolism / Q000494:pharmacology; D020024:Leukotriene Antagonists; D017997:Leukotriene C4 / Q000378:metabolism / Q000633:toxicity*; D017998:Leukotriene D4 / Q000378:metabolism / Q000633:toxicity*; D017999:Leukotriene E4 / Q000378:metabolism / Q000633:toxicity*; D008565:Membrane Proteins; D051379:Mice; D009924:Organ Culture Techniques; D010010:Osteoclasts / Q000187:drug effects*; D011869:Radioligand Assay; D051381:Rats; D018077:Receptors, Leukotriene; D014407:Tumor Cells, Cultured
chemical_list: D007191:Indazoles; D020024:Leukotriene Antagonists; D008565:Membrane Proteins; D018077:Receptors, Leukotriene; C054217:ICI 198615; D017997:Leukotriene C4; D017998:Leukotriene D4; D017999:Leukotriene E4; C412375:cysteinyl leukotriene receptor 2; C083169:leukotriene D4 receptor; D002118:Calcium