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Isolation of a cDNA sequence of rabbit GDF5 (mature form) and pattern of its mRNA expression during periosteal chondrogenesis
Authors:Arunik Sanyal  Gobinda Sarkar  James S Fitzsimmons  Shawn W O’Driscoll
Institution:Cartilage and Connective Tissue Research Laboratory, Department of Orthopedics, Mayo Clinic, 3-69 Medical Sciences Building, Rochester, MN 55905, USA.
Abstract:Articular cartilage has a limited ability for repair and/or regeneration. Periosteal grafts, having chondrogenic potential, are used clinically and in experimental models to study the repair and regeneration of cartilage. Growth/differentiation factor 5 (GDF5), recently shown to be involved in chondrogenesis and normal skeletal development, is a bioactive candidate for augmenting the repair of damaged cartilage. In order to investigate the role of GDF5 during periosteal chondrogenesis, the rabbit sequence must be known, as most experimental models involve rabbit tissues. For this purpose, the complete rabbit-specific cDNA sequence of the mature form of GDF5 was determined. Mature rabbit GDF5 was found to be 100% identical to that of human GDF5 at the amino acid level. Using the cDNA sequence, specific primers for PCR were designed. Quantitative RT-PCR, using rabbit-specific primers, showed up-regulation of GDF5 mRNAs early during periosteal chondrogenesis suggesting its potential involvement in this process. The timing and magnitude of this expression was markedly stimulated by TGF-beta 1, which has already been shown to be a potent inducer of periosteal chondrogenesis.
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