ADAMTS-1-knockout mice do not exhibit abnormalities in aggrecan turnover in vitro or in vivo.
OBJECTIVE
To determine the role of the proteinase ADAMTS-1 in normal and accelerated catabolism of aggrecan in articular and growth plate cartilage of mice.


METHODS
Expression of ADAMTS-1 was determined using reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of RNA isolated from microdissected chondrocytes from different zones of mouse growth plate and articular cartilage. Real-time RT-PCR for ADAMTS-4, ADAMTS-5, and ADAMTS-9 was performed on femoral head cartilage of wild-type (WT) and ADAMTS-1-knockout (KO) mice. Histologic and immunohistologic evaluation of growth plate and articular cartilage was performed in WT and KO mice from birth to 12 weeks of age. The effect of ADAMTS-1 ablation on cartilage proteoglycan loss was studied in antigen-induced arthritis (AIA). Aggrecan catabolism in WT and KO mice was studied in an in vitro model of cartilage degradation, by quantitation of glycosaminoglycan loss and histologic, immunohistologic, and Western immunoblot analyses.


RESULTS
ADAMTS-1 messenger RNA (mRNA) was expressed in normal mouse articular and growth plate cartilage and was up-regulated in terminal hypertrophic differentiation of growth plate chondrocytes. There was no difference in mRNA levels in the cartilage of WT compared with KO mice for the other potential aggrecanases ADAMTS-4, ADAMTS-5, or ADAMTS-9. ADAMTS-1-KO mice were significantly smaller than their WT littermates; however, no morphologic differences between the genotypes were evident in growth plate or articular cartilage from birth to skeletal maturity (12-16 weeks). Similarly, no difference in cartilage aggrecan content or presence of aggrecan degradation products was detected between WT and KO mice. There was no difference between WT and KO mice in the degree of synovial inflammation or depletion of cartilage aggrecan in AIA. There was no difference between WT and KO cartilage in either basal or stimulated aggrecan loss in vitro; however, subtle changes in the aggrecanase-generated aggrecan catabolites were observed in interleukin-1-treated cartilage.


CONCLUSIONS
Although ADAMTS-1 is expressed in articular and growth plate cartilage and is able to cleave aggrecan at physiologically relevant sites, our results indicate that it does not play a significant nonredundant role in normal cartilage and bone development and growth. Similarly, ablation of ADAMTS-1 offered no protection from accelerated aggrecanolysis in an inflammatory model of arthritis or in an in vitro model of early cartilage degradation. ADAMTS-1 does not appear to be a viable target for treatment of cartilage destruction in arthritis.
Little
Chris B
CB
Arthritis Research Group, University of Melbourne, Royal Children's Hospital, Parkville, Victoria, Australia. cblittle@med.usyd.edu.au
Mittaz
Laureane
L
Belluoccio
Daniele
D
Rogerson
Fraser M
FM
Campbell
Ian K
IK
Meeker
Clare T
CT
Bateman
John F
JF
Pritchard
Melanie A
MA
Fosang
Amanda J
AJ
D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; D053674:Aggrecans; D000818:Animals; D001168:Arthritis / Q000209:etiology; D001846:Bone Development; D002356:Cartilage / Q000378:metabolism*; D019483:Disintegrins / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis / Q000378:metabolism*; D006132:Growth Plate / Q000378:metabolism; D066298:In Vitro Techniques; D007596:Joints / Q000254:growth & development; D037181:Lectins, C-Type; D008666:Metalloendopeptidases / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D011509:Proteoglycans / Q000096:biosynthesis / Q000378:metabolism*
C508320:Acan protein, mouse; D053674:Aggrecans; D019483:Disintegrins; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D051722:ADAM Proteins; D000071097:ADAMTS1 Protein; C486323:Adamts1 protein, mouse; D008666:Metalloendopeptidases