title: Direct stimulation of osteoclastogenesis by MIP-1alpha: evidence obtained from studies using RAW264 cell clone highly responsive to RANKL.
abstract: Macrophage inflammatory protein-1alpha (MIP-1alpha) is a member of the CC chemokines. We have previously reported the use of a whole bone marrow culture system to show that MIP-1alpha stimulates the formation of osteoclast-like multinucleated cells. Here we use rat bone marrow cells deprived of stromal cells, and clones obtained from murine macrophage-like cell line RAW264 to show that MIP-1alpha acts directly on cells in osteoclast lineage. We obtained several types of RAW264 cell clones, one of these clones, designated as RAW264 cell D clone (D clone), showed an extremely high response to receptor activator of NFkappaB ligand (RANKL) and tumor necrosis factor-alpha (TNF-alpha), while the other clone, RAW264 cell N clone (N clone), demonstrated no response to RANKL or TNF-alpha. Although both clones expressed receptor activator NFkappaB (RANK) before being stimulated for differentiation, only the D clone expressed cathepsin K when cells were stimulated to differentiate to osteoclasts. MIP-1alpha stimulated the formation of mononuclear preosteoclast-like cells from rat bone marrow cells deprived of stromal cells. MIP-1alpha also stimulated formation of osteoclast-like multinucleated cells from the D clone, when these cells were stimulated with RANKL and TNF-alpha. These findings provide strong evidence to show that MIP-1alpha acts directly on cells in the osteoclast lineage to stimulate osteoclastogenesis. Furthermore, pretreatment of RAW264 cell D clone with MIP-1alpha significantly induced adhesion properties of these cells to primary osteoblasts, suggesting a crucial role for MIP-1alpha in the regulation of the interaction between osteoclast precursors and osteoblasts in osteoclastogenesis.
authors.affiliation: Oral Cellular and Molecular Biology, Division of Oral Biological Sciences, Faculty of Dental Science, Kyushu University, Fukuoka, Fukuoka 812-8582, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Watanabe
authors.forename: Toshio
authors.initials: T
authors.lastname: Kukita
authors.forename: Akiko
authors.initials: A
authors.lastname: Kukita
authors.forename: Naohisa
authors.initials: N
authors.lastname: Wada
authors.forename: Kazuko
authors.initials: K
authors.lastname: Toh
authors.forename: Kengo
authors.initials: K
authors.lastname: Nagata
authors.forename: Hisayuki
authors.initials: H
authors.lastname: Nomiyama
authors.forename: Tadahiko
authors.initials: T
authors.lastname: Iijima
mesh_terms: D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells; D002352:Carrier Proteins / Q000494:pharmacology; D056657:Cathepsin K; D002403:Cathepsins / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion; D002454:Cell Differentiation; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D002999:Clone Cells; D004305:Dose-Response Relationship, Drug; D007527:Isoenzymes / Q000378:metabolism; D019402:Macrophage Inflammatory Proteins / Q000494:pharmacology*; D008297:Male; D008562:Membrane Glycoproteins / Q000494:pharmacology; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects; D010010:Osteoclasts / Q000166:cytology* / Q000187:drug effects; D010012:Osteogenesis / Q000187:drug effects*; D053245:RANK Ligand; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D053246:Receptor Activator of Nuclear Factor-kappa B; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013268:Stimulation, Chemical; D000071681:Tartrate-Resistant Acid Phosphatase; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D002352:Carrier Proteins; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D007527:Isoenzymes; D019402:Macrophage Inflammatory Proteins; D008562:Membrane Glycoproteins; D053245:RANK Ligand; D012333:RNA, Messenger; D053246:Receptor Activator of Nuclear Factor-kappa B; C505687:Tnfrsf11a protein, mouse; C505689:Tnfsf11 protein, mouse; D014409:Tumor Necrosis Factor-alpha; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; D002403:Cathepsins; D056657:Cathepsin K; C534132:Ctsk protein, mouse; C534133:Ctsk protein, rat