title: MED and PSACH COMP mutations affect chondrogenesis in chicken limb bud micromass cultures.
abstract: Mutations in cartilage oligomeric matrix protein (COMP) cause pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED). We studied the effects of over-expression of wild type and mutant COMP on early stages of chondrogenesis in chicken limb bud micromass cultures. Cells were transduced with RCAS virus harboring wild type or mutant (C328R, PSACH; T585R, MED) COMP cDNAs and cultured for 3, 4, and 5 days. The effect of COMP constructs on chondrogenesis was assessed by analyzing mRNA and protein expression of several COMP binding partners. Cell viability was assayed, and evaluation of apoptosis was performed by monitoring caspase 3 processing. Over-expression of COMP, and especially expression of COMP mutants, had a profound affect on the expression of syndecan 3 and tenascin C, early markers of chondrogenesis. Over-expression of COMP did not affect levels of type II collagen or matrilin-3; however, there were increases in type IX collagen expression and sulfated proteoglycan synthesis, particularly at day 5 of harvest. In contrast to cells over-expressing COMP, cells with mutant COMP showed reduction in type IX collagen expression and increased matrilin 3 expression. Finally, reduction in cell viability, and increased activity of caspase 3, at days 4 and 5, were observed in cultures expressing either wild type or mutant COMP. MED, and PSACH mutations, despite displaying phenotypic differences, demonstrated only subtle differences in their cellular viability and mRNA and protein expression of components of the extracellular matrix, including those that interact with COMP. These results suggest that COMP mutations, by disrupting normal interactions between COMP and its binding partners, significantly affect chondrogenesis.
authors.affiliation: Division of Rheumatology, Department of Medicine, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
authors.forename: J
authors.initials: J
authors.lastname: Roman-Blas
authors.forename: A S
authors.initials: AS
authors.lastname: Dion
authors.forename: M R
authors.initials: MR
authors.lastname: Seghatoleslami
authors.forename: K
authors.initials: K
authors.lastname: Giunta
authors.forename: P
authors.initials: P
authors.lastname: Oca
authors.forename: S A
authors.initials: SA
authors.lastname: Jimenez
authors.forename: C J
authors.initials: CJ
authors.lastname: Williams
mesh_terms: D000130:Achondroplasia / Q000235:genetics* / Q000473:pathology; D000595:Amino Acid Sequence; D000818:Animals; D064236:Cartilage Oligomeric Matrix Protein; D018929:Cell Culture Techniques; D002470:Cell Survival; D002478:Cells, Cultured; D002645:Chickens; D020219:Chondrogenesis / Q000502:physiology*; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000235:genetics* / Q000378:metabolism; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D018878:Limb Buds / Q000166:cytology / Q000502:physiology*; D064235:Matrilin Proteins; D008969:Molecular Sequence Data; D009154:Mutation; D010009:Osteochondrodysplasias / Q000235:genetics* / Q000473:pathology; D016415:Sequence Alignment; D053667:Syndecans / Q000235:genetics / Q000378:metabolism; D019063:Tenascin / Q000235:genetics / Q000378:metabolism
chemical_list: D064236:Cartilage Oligomeric Matrix Protein; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D064235:Matrilin Proteins; D053667:Syndecans; C577553:TSP5 protein, human; D019063:Tenascin