Selective assembly and remodelling of collagens II and IX associated with expression of the chondrocyte hypertrophic phenotype.
The assembly and resorption of the extracellular matrix in the physis of the growth plate are poorly understood. By examining isolated fetal growth plate chondrocytes in culture and using immunochemical methods we show that type II collagen, proteoglycan aggrecan, and type IX collagen are assembled into a matrix that is initially enriched in type II collagen over proteoglycan and type IX collagen. When compared to the content of the COL2 domain in the alpha(1)(IX) chain it is evident that the majority ( 90%) of type IX molecules lack the NC4 domain unlike in articular cartilage. During matrix assembly the molar ratio of type II/COL2 of alpha(1)(IX) varied from 25:1 to 2.5:1. Following expression of the hypertrophic phenotype (initiation of type X collagen synthesis) there are parallel changes in both collagen and proteoglycan contents (inversely related to collagenase cleavage of type II collagen). The NC4 domain is then selectively, rapidly and irreversibly removed as mineralization is initiated, leaving the alpha(1)(IX) chain COL2 domain. Subsequently as mineralization progresses type II and type IX collagen (COL2 domain), but not the proteoglycan aggrecan, are resorbed coincident with a markedly increased cleavage of type II collagen by collagenase as mineral is deposited in the matrix. This study, therefore reveals a carefully orchestrated series of events in matrix assembly and resorption that prepares the extracellular matrix for mineralization.
Mwale
F
F
Joint Diseases Laboratory, Shriners Hospitals for Children and Division of Surgical Research, Department of Surgery, McGill University, Montreal, Quebec, Canada.
Billinghurst
C
C
Wu
W
W
Alini
M
M
Webber
C
C
Reiner
A
A
Ionescu
M
M
Poole
J
J
Poole
A R
AR
D000818:Animals; D015153:Blotting, Western; D002356:Cartilage / Q000196:embryology; D002417:Cattle; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism*; D003094:Collagen / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D004306:Dose-Response Relationship, Immunologic; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006025:Glycosaminoglycans / Q000378:metabolism; D006984:Hypertrophy / Q000378:metabolism*; D007118:Immunoassay; D008954:Models, Biological; D010455:Peptides / Q000378:metabolism; D010641:Phenotype; D017434:Protein Structure, Tertiary; D013997:Time Factors
D000939:Epitopes; D006025:Glycosaminoglycans; D010455:Peptides; D003094:Collagen