title: Analysis of CCN4 Function in Osteogenic and Osteoclastic Cells Using Gain and Loss of Function Approaches.
abstract: Analysis of CCN4 function in bone was assessed using both gain and loss of function approaches. In mice this was done by genetic engineering and homologous recombination to create mice with complete ablation of the protein. For human skeletal cells adenoviral gene transfer and shRNA were used for gain and loss of function respectively. Here we describe procedures used to make and then analyze osteogenic and osteoclastic cells with or without CCN4 to determine its role in osteogenic differentiation.
authors.affiliation: Center for Promotion of Dental Education and International Collaboration, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama City, Okayama, 700-8558, Japan.
authors.forename: Azusa
authors.initials: A
authors.lastname: Maeda
authors.affiliation: Molecular Biology of Bones and Teeth Section, Craniofacial and Skeletal Diseases Branch, National Institutes of Health, Bethesda, MD, 20892, USA. myoung@dir.nidcr.nih.gov.
authors.forename: Marian
authors.initials: M
authors.lastname: Young
authors.affiliation: Department of Molecular Biology and Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama City, Okayama, 700-8558, Japan.
authors.forename: Mitsuaki
authors.initials: M
authors.lastname: Ono
mesh_terms: D000818:Animals; D001862:Bone Resorption / Q000235:genetics / Q000378:metabolism; D055511:CCN Intercellular Signaling Proteins / Q000235:genetics* / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D000073659:Gain of Function Mutation; D060005:Genotyping Techniques; D006801:Humans; D000073658:Loss of Function Mutation; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D010010:Osteoclasts / Q000378:metabolism*; D010012:Osteogenesis / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism*
chemical_list: D055511:CCN Intercellular Signaling Proteins; C492067:CCN4 protein, mouse; D011518:Proto-Oncogene Proteins
keywords: CCN4; Gene knockout; Osteogenic differentiation; Osteoprogenitor cell