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Article ; Online: Isolation and culture of primary chicken osteoclasts.

Collin-Osdoby, Patricia / Osdoby, Philip

Methods in molecular biology (Clifton, N.J.)

2012 Volume 816, Page(s) 119–143

Abstract: Osteoclasts originate from hematopoietic myeloid progenitors that differentiate into specialized multinucleated cells uniquely capable of resorbing bone in both physiological and pathological conditions. Osteoclast numbers and degradative activities ... ...

Abstract	Osteoclasts originate from hematopoietic myeloid progenitors that differentiate into specialized multinucleated cells uniquely capable of resorbing bone in both physiological and pathological conditions. Osteoclast numbers and degradative activities increase in various inflammatory disorders of bone and certain bone oncologies, thereby causing bone loss that may weaken the skeleton, increase fracture incidence, and disturb marrow function. Many valuable insights have been obtained through the use of osteoclasts directly isolated from the bones of chickens fed a low calcium diet to enhance osteoclastogenesis and bone resorption. Particular advantages of this system include the abundance and highly resorptive nature of isolated chicken osteoclasts compared with those directly obtained from other species. After enzymatic release from the harvested bones, osteoclasts may be partially purified by density gradient sedimentation, bone substrate attachment, and/or immunomagnetic capture. Thereafter, osteoclast preparations may be analyzed, either directly or following some period of culture, to investigate their properties (biochemical, immunological, molecular, cell biological), resorptive function, and modulatory responses to various stimuli. Here, we present common procedures for the isolation, culture, and general study of chicken osteoclasts.
MeSH term(s)	Animals ; Antigens/analysis ; Bone Resorption ; Bone and Bones/cytology ; Cell Culture Techniques/methods ; Cell Movement ; Cell Separation/methods ; Cells, Cultured ; Chickens ; Histocytochemistry/methods ; Osteoclasts/cytology ; Osteoclasts/immunology ; Osteoclasts/ultrastructure ; Staining and Labeling/methods ; Tissue Fixation/methods
Chemical Substances	Antigens
Language	English
Publishing date	2012
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-61779-415-5_9
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Article ; Online: RANKL-mediated osteoclast formation from murine RAW 264.7 cells.

Collin-Osdoby, Patricia / Osdoby, Philip

Methods in molecular biology (Clifton, N.J.)

2012 Volume 816, Page(s) 187–202

Abstract: Extensive research efforts over the years have provided us with great insights into how bone-resorbing osteoclasts (OCs) develop and function and, based on such work, valuable antiresorptive therapies have been developed to help combat the excessive bone ...

Abstract	Extensive research efforts over the years have provided us with great insights into how bone-resorbing osteoclasts (OCs) develop and function and, based on such work, valuable antiresorptive therapies have been developed to help combat the excessive bone loss that occurs in numerous skeletal disorders. The RAW 264.7 murine cell line has proven to be an important tool for in vitro studies of OC formation and function, having particular advantages over the use of OCs generated from primary bone marrow cell populations or directly isolated from murine bones. These include their ready access and availability, simple culture for this pure macrophage/pre-OC population, sensitive and rapid development into highly bone-resorptive OCs expressing hallmark OC characteristics following their RANKL stimulation, abundance of RAW cell-derived OCs that can be generated to provide large amounts of study material, relative ease of transfection for genetic and regulatory manipulation, and close correlation in characteristics, gene expression, signaling, and developmental or functional processes between RAW cell-derived OCs and OCs either directly isolated from murine bones or formed in vitro from primary bone marrow precursor cells. Here, we describe methods for the culture and RANKL-mediated differentiation of RAW cells into bone-resorptive OCs as well as procedures for their enrichment, characterization, and general use in diverse analytical assays.
MeSH term(s)	Animals ; Bone Resorption ; Bone and Bones/cytology ; Cell Culture Techniques/methods ; Cell Differentiation ; Cell Line ; Macrophages/cytology ; Macrophages/metabolism ; Mice ; Osteoclasts/cytology ; Osteoclasts/metabolism ; RANK Ligand/metabolism
Chemical Substances	RANK Ligand
Language	English
Publishing date	2012
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-61779-415-5_13
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Article ; Online: Regulation of vascular calcification by osteoclast regulatory factors RANKL and osteoprotegerin.

Collin-Osdoby, Patricia

Circulation research

2004 Volume 95, Issue 11, Page(s) 1046–1057

Abstract: Vascular calcification often occurs with advancing age, atherosclerosis, various metabolic disorders such as diabetes mellitus and end-stage renal disease, or in rare genetic diseases, leading to serious clinical consequences. Such mineralization can ... ...

Abstract	Vascular calcification often occurs with advancing age, atherosclerosis, various metabolic disorders such as diabetes mellitus and end-stage renal disease, or in rare genetic diseases, leading to serious clinical consequences. Such mineralization can occur at various sites (cardiac valves, arterial intima or media, capillaries), involve localized or diffuse widespread calcification, and result from numerous causes that provoke active inflammatory and osteogenic processes or disordered mineral homeostasis. Although valuable research has defined many key factors and cell types involved, surprising new insights continue to arise that deepen our understanding and suggest novel research directions or strategies for clinical intervention in calcific vasculopathies. One emerging area in vascular biology involves the RANKL/RANK/OPG system, molecules of the tumor necrosis factor-related family recently discovered to be critical regulators of immune and skeletal biology. Evidence is accumulating that such signals may be expressed, regulated, and function in vascular physiology and pathology in unique ways to promote endothelial cell survival, angiogenesis, monocyte or endothelial cell recruitment, and smooth muscle cell osteogenesis and calcification. Concerted research efforts are greatly needed to understand these potential roles, clarify whether RANKL (receptor activator of nuclear factor kappaB ligand) promotes and osteoprotegerin (OPG) protects against vascular calcification, define how OPG genetic polymorphisms relate to cardiovascular disease, and learn whether elevated serum OPG levels reflect endothelial dysfunction in patients. Overall, the RANKL/RANK/OPG system may mediate important and complex links between the vascular, skeletal, and immune systems. Thus, these molecules may play a central role in regulating the development of vascular calcification coincident with declines in skeletal mineralization with age, osteoporosis, or disease.
MeSH term(s)	Animals ; Arteriosclerosis/physiopathology ; Bone and Bones/physiology ; Calcinosis/complications ; Calcinosis/physiopathology ; Carrier Proteins/pharmacology ; Carrier Proteins/physiology ; Endothelium, Vascular/drug effects ; Glycoproteins/pharmacology ; Glycoproteins/physiology ; Humans ; Immune System/physiology ; Membrane Glycoproteins/pharmacology ; Membrane Glycoproteins/physiology ; Mice ; Mice, Knockout ; Muscle, Smooth, Vascular/drug effects ; Osteoclasts/physiology ; Osteoporosis/complications ; Osteoporosis/physiopathology ; Osteoprotegerin ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; Receptors, Cytoplasmic and Nuclear/physiology ; Receptors, Tumor Necrosis Factor ; Vascular Diseases/complications ; Vascular Diseases/physiopathology
Chemical Substances	Carrier Proteins ; Glycoproteins ; Membrane Glycoproteins ; Osteoprotegerin ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; Receptors, Cytoplasmic and Nuclear ; Receptors, Tumor Necrosis Factor ; TNFRSF11A protein, human ; TNFRSF11B protein, human ; TNFSF11 protein, human ; Tnfrsf11a protein, mouse ; Tnfrsf11b protein, mouse ; Tnfsf11 protein, mouse
Language	English
Publishing date	2004-11-26
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, P.H.S. ; Review
ZDB-ID	80100-8
ISSN	1524-4571 ; 0009-7330 ; 0931-6876
ISSN (online)	1524-4571
ISSN	0009-7330 ; 0931-6876
DOI	10.1161/01.RES.0000149165.99974.12
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Article: Primary isolation and culture of chicken osteoclasts.

Collin-Osdoby, Patricia / Anderson, Fred / Osdoby, Philip

Methods in molecular medicine

2003 Volume 80, Page(s) 65–88

MeSH term(s)	Animals ; Antigens/analysis ; Antigens/immunology ; Bone Resorption/metabolism ; Calcium/deficiency ; Cell Culture Techniques/methods ; Cell Movement/physiology ; Cell Separation/methods ; Chickens ; Clinical Laboratory Techniques/methods ; Immunomagnetic Separation/methods ; Osteoclasts/cytology ; Osteoclasts/immunology ; Osteoclasts/metabolism ; Staining and Labeling/methods
Chemical Substances	Antigens ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2003
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S.
ISSN	1543-1894
ISSN	1543-1894
DOI	10.1385/1-59259-366-6:65
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Article: Vascular biology and the skeleton.

Brandi, Maria Luisa / Collin-Osdoby, Patricia

Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

2006 Volume 21, Issue 2, Page(s) 183–192

MeSH term(s)	Bone Development ; Bone Diseases/pathology ; Bone Diseases/physiopathology ; Bone and Bones/blood supply ; Bone and Bones/cytology ; Endothelium, Vascular/physiology ; Endothelium, Vascular/physiopathology ; Humans
Language	English
Publishing date	2006-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	632783-7
ISSN	1523-4681 ; 0884-0431
ISSN (online)	1523-4681
ISSN	0884-0431
DOI	10.1359/JBMR.050917
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Article: SDF-1 increases recruitment of osteoclast precursors by upregulation of matrix metalloproteinase-9 activity.

Yu, Xuefeng / Collin-Osdoby, Patricia / Osdoby, Philip

Connective tissue research

2003 Volume 44 Suppl 1, Page(s) 79–84

Abstract: Although chemokines play essential roles in the trafficking and homing of many circulating hematopoietic cell types, their potential influences on osteoclast (OC) recruitment or bone remodeling are not well known. Therefore, chemokine receptor expression ...

Abstract	Although chemokines play essential roles in the trafficking and homing of many circulating hematopoietic cell types, their potential influences on osteoclast (OC) recruitment or bone remodeling are not well known. Therefore, chemokine receptor expression was analyzed by RNase protection assay during OC formation induced by RANKL in a murine mononuclear cell line (RAW 264.7). Relatively high CXCR4 expression was detected in RAW cells (pre-OCs), whereas CXCR4 levels were downregulated during RAW-OC development. SDF-1, the unique ligand for CXCR4, stimulated RAW cell production of matrix metalloproteinase (MMP)-9 activity, a matrix-degrading enzyme essential for pre-OC migration into the developing bone marrow cavity. Induced MMP-9 activity in RAW cells was associated with their increased MMP-dependent transmigration through a collagen gel in response to SDF-1. We conclude that SDF-1 stimulation of MMP-9 activity in pre-OCs may be a key aspect of their recruitment to bone and migration within the marrow to sites for OC differentiation and bone resorption.
MeSH term(s)	Animals ; Carrier Proteins/pharmacology ; Cell Differentiation/drug effects ; Cell Line ; Cell Movement/drug effects ; Chemokine CXCL12 ; Chemokines, CXC/pharmacology ; Macrophages ; Matrix Metalloproteinase 9/biosynthesis ; Membrane Glycoproteins/pharmacology ; Mice ; Osteoclasts/cytology ; Osteoclasts/drug effects ; Osteoclasts/enzymology ; RANK Ligand ; RNA, Messenger/metabolism ; Receptor Activator of Nuclear Factor-kappa B ; Receptors, CXCR4/biosynthesis ; Receptors, CXCR4/genetics ; Stem Cells/cytology ; Stem Cells/drug effects ; Stem Cells/enzymology ; Up-Regulation
Chemical Substances	Carrier Proteins ; Chemokine CXCL12 ; Chemokines, CXC ; Cxcl12 protein, mouse ; Membrane Glycoproteins ; RANK Ligand ; RNA, Messenger ; Receptor Activator of Nuclear Factor-kappa B ; Receptors, CXCR4 ; Tnfrsf11a protein, mouse ; Tnfsf11 protein, mouse ; Matrix Metalloproteinase 9 (EC 3.4.24.35)
Language	English
Publishing date	2003
Publishing country	England
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	185551-7
ISSN	1607-8438 ; 0300-8207 ; 0091-1690
ISSN (online)	1607-8438
ISSN	0300-8207 ; 0091-1690
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Article: RANKL stimulates inducible nitric-oxide synthase expression and nitric oxide production in developing osteoclasts. An autocrine negative feedback mechanism triggered by RANKL-induced interferon-beta via NF-kappaB that restrains osteoclastogenesis and bone resorption.

Zheng, Hong / Yu, Xuefeng / Collin-Osdoby, Patricia / Osdoby, Philip

The Journal of biological chemistry

2006 Volume 281, Issue 23, Page(s) 15809–15820

Abstract: Nitric oxide (NO) is a multifunctional signaling molecule and a key vasculoprotective and potential osteoprotective factor. NO regulates normal bone remodeling and pathological bone loss in part through affecting the recruitment, formation, and activity ... ...

Abstract	Nitric oxide (NO) is a multifunctional signaling molecule and a key vasculoprotective and potential osteoprotective factor. NO regulates normal bone remodeling and pathological bone loss in part through affecting the recruitment, formation, and activity of bone-resorbing osteoclasts. Using murine RAW 264.7 and primary bone marrow cells or osteoclasts formed from them by receptor activator of NF-kappaB ligand (RANKL) differentiation, we found that inducible nitric-oxide synthase (iNOS) expression and NO generation were stimulated by interferon (IFN)-gamma or lipopolysaccharide, but not by interleukin-1 or tumor necrosis factor-alpha. Surprisingly, iNOS expression and NO release were also triggered by RANKL. This response was time- and dose-dependent, required NF-kappaB activation and new protein synthesis, and was specifically blocked by the RANKL decoy receptor osteoprotegerin. Preventing RANKL-induced NO (via iNOS-selective inhibition or use of marrow cells from iNOS-/- mice) increased osteoclast formation and bone pit resorption, indicating that such NO normally restrains RANKL-mediated osteoclastogenesis. Additional studies suggested that RANKL-induced NO inhibition of osteoclast formation does not occur via NO activation of a cGMP pathway. Because IFN-beta is also a RANKL-induced autocrine negative feedback inhibitor that limits osteoclastogenesis, we investigated whether IFN-beta is involved in this novel RANKL/iNOS/NO autoregulatory pathway. IFN-beta was induced by RANKL and stimulated iNOS expression and NO release, and a neutralizing antibody to IFN-beta inhibited iNOS/NO elevation in response to RANKL, thereby enhancing osteoclast formation. Thus, RANKL-induced IFN-beta triggers iNOS/NO as an important negative feedback signal during osteoclastogenesis. Specifically targeting this novel autoregulatory pathway may provide new therapeutic approaches to combat various osteolytic bone diseases.
MeSH term(s)	Animals ; Base Sequence ; Bone Resorption ; Carrier Proteins/physiology ; Cell Line ; DNA Primers ; Enzyme Inhibitors/pharmacology ; Interferon-beta/biosynthesis ; Interferon-beta/physiology ; Membrane Glycoproteins/physiology ; Mice ; NF-kappa B/metabolism ; Nitric Oxide/biosynthesis ; Nitric Oxide Synthase Type II/antagonists & inhibitors ; Nitric Oxide Synthase Type II/metabolism ; Osteoclasts/enzymology ; Osteoclasts/metabolism ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; Reverse Transcriptase Polymerase Chain Reaction
Chemical Substances	Carrier Proteins ; DNA Primers ; Enzyme Inhibitors ; Membrane Glycoproteins ; NF-kappa B ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; Tnfrsf11a protein, mouse ; Tnfsf11 protein, mouse ; Nitric Oxide (31C4KY9ESH) ; Interferon-beta (77238-31-4) ; Nitric Oxide Synthase Type II (EC 1.14.13.39)
Language	English
Publishing date	2006-04-12
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.M513225200
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Article: CCR1 chemokines promote the chemotactic recruitment, RANKL development, and motility of osteoclasts and are induced by inflammatory cytokines in osteoblasts.

Yu, Xuefeng / Huang, Yuefang / Collin-Osdoby, Patricia / Osdoby, Philip

Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

2004 Volume 19, Issue 12, Page(s) 2065–2077

Abstract: Unlabelled: Chemoattractants that recruit OC precursors to locally inflamed sites of resorption are not well known. A chemokine receptor, CCR1, was expressed in OC precursors and elevated in mature OCs, and its ligands promoted OC precursor recruitment, ...

Abstract	Unlabelled: Chemoattractants that recruit OC precursors to locally inflamed sites of resorption are not well known. A chemokine receptor, CCR1, was expressed in OC precursors and elevated in mature OCs, and its ligands promoted OC precursor recruitment, RANKL development, and OC motility. Cytokines induced OB release of such chemokines, which may therefore significantly contribute to inflammatory bone loss. Introduction: Chemokines, primarily of two major (CXC, CC) families, are essential signals for the trafficking and localization of circulating hematopoietic cells into tissues. However, little is known about their potential roles in osteoclast (OC) recruitment, development, or function. Previously, we analyzed CXC receptors in murine OC precursors and found high expression of CXCR4 that mediated their stromal-derived factor-1(SDF-1)-induced chemotaxis and collagen invasion. Here, we investigated if CC receptors and ligands, which are elevated in inflammatory and other osteolytic diseases, also play important roles in the recruitment, formation, or activity of murine bone-resorptive OCs. Materials and methods: CC chemokine receptor (CCR) mRNA expression was analyzed during OC formation induced by RANKL in murine RAW 264.7 cells and primary marrow cells. Corresponding CC chemokines were tested for their ability to elicit precursor chemotaxis or OC development, or to influence motility, bone resorption, adhesion, or survival in RANKL-differentiated OCs. Constitutive and inflammatory cytokine-induced release of the chemokines macrophage inflammatory protein-1alpha (MIP-1alpha) and regulated on activation, normal T-cell expressed and secreted (RANTES) was measured by ELISA for OCs, osteoblasts (OBs), and their precursor cells. Results: CCR1 was expressed in murine marrow cells, the most prominent CCR in RAW cells, and upregulated by RANKL in marrow or RAW cells. Chemokines that bind CCR1 (MIP-1alpha, RANTES, and monocyte chemoattractant protein-3 [MCP-3]) were produced to varying degrees by murine OCs, OBs, and their precursors, and markedly increased by interleukin (IL)-1alpha and TNFalpha in differentiating OBs. RANTES, and especially MIP-1alpha, increased mature OC motility, but did not alter OC resorption activity, adhesion, or survival. All three chemokines stimulated chemotaxis of marrow or RAW cell precursors, leading to the greater formation of OCs (in number and size) after RANKL development of such chemoattracted marrow cells. All three chemokines also directly and dramatically enhanced OC formation in marrow cultures, through a pathway dependent on the presence of RANKL but without altering RANK expression. Conclusions: Pathological increases in secretion of these chemokines from activated OBs or other cells may potently stimulate the chemotactic recruitment and RANKL formation of bone-resorptive OCs, thereby exacerbating local osteolysis in multiple skeletal diseases.
MeSH term(s)	Animals ; Bone Diseases/metabolism ; Bone Resorption ; Bone and Bones/metabolism ; Carrier Proteins/metabolism ; Cell Movement ; Cells, Cultured ; Centrifugation, Density Gradient ; Chemokine CCL3 ; Chemokine CCL4 ; Chemokine CCL5/metabolism ; Chemokine CCL7 ; Chemokine CXCL12 ; Chemokines/metabolism ; Chemokines, CXC/metabolism ; Chemotaxis ; Cytokines/metabolism ; Dose-Response Relationship, Drug ; Enzyme-Linked Immunosorbent Assay ; Inflammation ; Ligands ; Macrophage Inflammatory Proteins/metabolism ; Membrane Glycoproteins/metabolism ; Mice ; Models, Biological ; Monocyte Chemoattractant Proteins/metabolism ; Osteoblasts/metabolism ; Osteoclasts/metabolism ; RANK Ligand ; RNA/chemistry ; RNA, Messenger/metabolism ; Receptor Activator of Nuclear Factor-kappa B ; Receptors, CCR1 ; Receptors, Chemokine/metabolism ; Receptors, Chemokine/physiology ; Ribonucleases/chemistry ; Up-Regulation
Chemical Substances	Carrier Proteins ; Ccl7 protein, mouse ; Ccr1 protein, mouse ; Chemokine CCL3 ; Chemokine CCL4 ; Chemokine CCL5 ; Chemokine CCL7 ; Chemokine CXCL12 ; Chemokines ; Chemokines, CXC ; Cxcl12 protein, mouse ; Cytokines ; Ligands ; Macrophage Inflammatory Proteins ; Membrane Glycoproteins ; Monocyte Chemoattractant Proteins ; RANK Ligand ; RNA, Messenger ; Receptor Activator of Nuclear Factor-kappa B ; Receptors, CCR1 ; Receptors, Chemokine ; Tnfrsf11a protein, mouse ; Tnfsf11 protein, mouse ; RNA (63231-63-0) ; Ribonucleases (EC 3.1.-)
Language	English
Publishing date	2004-12
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	632783-7
ISSN	1523-4681 ; 0884-0431
ISSN (online)	1523-4681
ISSN	0884-0431
DOI	10.1359/JBMR.040910
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Article: RANKL-mediated osteoclast formation from murine RAW 264.7 cells.

Collin-Osdoby, Patricia / Yu, Xuefeng / Zheng, Hong / Osdoby, Philip

Methods in molecular medicine

2003 Volume 80, Page(s) 153–166

MeSH term(s)	Animals ; Biological Assay/methods ; Bone Resorption ; Carrier Proteins/physiology ; Cell Culture Techniques/methods ; Cell Differentiation/physiology ; Clinical Laboratory Techniques/methods ; Membrane Glycoproteins/physiology ; Mice ; Osteoclasts/physiology ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B
Chemical Substances	Carrier Proteins ; Membrane Glycoproteins ; RANK Ligand ; Receptor Activator of Nuclear Factor-kappa B ; Tnfrsf11a protein, mouse ; Tnfsf11 protein, mouse
Language	English
Publishing date	2003
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S.
ISSN	1543-1894
ISSN	1543-1894
DOI	10.1385/1-59259-366-6:153
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Article: Stromal cell-derived factor-1 (SDF-1) recruits osteoclast precursors by inducing chemotaxis, matrix metalloproteinase-9 (MMP-9) activity, and collagen transmigration.

Yu, Xuefeng / Huang, Yuefang / Collin-Osdoby, Patricia / Osdoby, Philip

Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research

2003 Volume 18, Issue 8, Page(s) 1404–1418

Abstract: Unlabelled: Signals targeting OCs to bone and resorption sites are not well characterized. A chemoattractant receptor (CXCR4), highly expressed in murine OC precursors, mediated their chemokine (SDF-1)-induced chemoattraction, collagen transmigration, ... ...

Abstract	Unlabelled: Signals targeting OCs to bone and resorption sites are not well characterized. A chemoattractant receptor (CXCR4), highly expressed in murine OC precursors, mediated their chemokine (SDF-1)-induced chemoattraction, collagen transmigration, and MMP-9 expression. Thus, bone vascular and stromal SDF-1 may direct OC precursors into bone and marrow sites for development and bone resorption. Introduction: Although chemokines are essential for trafficking and homing of circulating hematopoietic cells under normal and pathological conditions, their potential roles in osteoclast (OC) recruitment or function are generally unknown. CXCR4 and its unique ligand, stromal cell-derived factor-1 (SDF-1), critically control the matrix metalloproteinase (MMP)-dependent targeting of hematopoietic cells into bone and within the marrow microenvironment. Therefore, SDF-1/CXCR4 may regulate OC precursor recruitment to sites for development and activation. Methods: Chemokine receptor mRNA expression was analyzed during OC formation induced by RANKL in murine RAW 264.7 cells. SDF-1 versus RANKL effects on chemotaxis, transcollagen migration, MMP-9 expression and activity, OC development, and bone resorption were evaluated in RAW cells or RAW-OCs. Results: CXCR4 was highly expressed in RAW cells and downregulated during their RANKL development into bone-resorptive RAW-OCs. SDF-1, but not RANKL, elicited RAW cell chemotaxis. Conversely, RANKL, but not SDF-1, promoted RAW-OC development, TRAP activity, cathepsin K expression, and bone pit resorption, and SDF-1 did not modify these RANKL responses. Both SDF-1 and RANKL increased MMP-9, a matrix-degrading enzyme essential for OC precursor migration into developing bone marrow cavities, and increased transcollagen migration of RAW cells in a MMP-dependent manner. SDF-1 also upregulated MMP-9 in various primary murine OC precursor cells. Because RANKL induced a higher, more sustained expression of MMP-9 in RAW cells than did SDF-1, MMP-9 may have an additional role in mature OCs. Consistent with this, MMP-9 upregulation during RANKL-induced RAW-OC development was necessary for initiation of bone pit resorption. Conclusions: SDF-1, a chemokine highly expressed by bone vascular endothelial and marrow stromal cells, may be a key signal for the selective attraction of circulating OC precursors into bone and their migration within marrow to appropriate perivascular stromal sites for RANKL differentiation into resorptive OCs. Thus, SDF-1 and RANKL likely serve complementary physiological functions, partly mediated through increases in MMP-9, to coordinate stages of OC precursor recruitment, development, and function.
MeSH term(s)	Animals ; Bone Resorption ; Carrier Proteins/pharmacology ; Chemokine CXCL12 ; Chemokines, CXC/pharmacology ; Chemotaxis/drug effects ; Collagen/metabolism ; Enzyme Induction/drug effects ; Matrix Metalloproteinase 2/metabolism ; Matrix Metalloproteinase 9/metabolism ; Membrane Glycoproteins/pharmacology ; Mice ; Osteoclasts/cytology ; Osteoclasts/drug effects ; Osteoclasts/enzymology ; Osteoclasts/metabolism ; RANK Ligand ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Receptor Activator of Nuclear Factor-kappa B ; Receptors, Chemokine/genetics ; Stem Cells/cytology ; Stem Cells/drug effects ; Stem Cells/enzymology ; Stem Cells/metabolism
Chemical Substances	Carrier Proteins ; Chemokine CXCL12 ; Chemokines, CXC ; Cxcl12 protein, mouse ; Membrane Glycoproteins ; RANK Ligand ; RNA, Messenger ; Receptor Activator of Nuclear Factor-kappa B ; Receptors, Chemokine ; Tnfrsf11a protein, mouse ; Tnfsf11 protein, mouse ; Collagen (9007-34-5) ; Matrix Metalloproteinase 2 (EC 3.4.24.24) ; Matrix Metalloproteinase 9 (EC 3.4.24.35)
Language	English
Publishing date	2003-08
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	632783-7
ISSN	1523-4681 ; 0884-0431
ISSN (online)	1523-4681
ISSN	0884-0431
DOI	10.1359/jbmr.2003.18.8.1404
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In stock of ZB MED Cologne/Königswinter

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In stock of ZB MED Cologne/Königswinter

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