ADAM-10 protein is present in human articular cartilage primarily in the membrane-bound form and is upregulated in osteoarthritis and in response to IL-1alpha in bovine nasal cartilage.
The objective of our study was to determine the tissue distribution and localization of ADAM-10 protein in human and bovine cartilage and the changes it undergoes with cartilage degeneration seen in osteoarthritis (OA) and under the influence of interleukin-1 (IL-1). Human normal and OA articular cartilage and bovine nasal cartilage cultured in the presence of IL-1alpha were processed for histology and immunohistochemistry. ADAM-10 protein was extracted from human cartilage and analyzed by Western blotting using anti-ADAM-10 antibodies. Fluor S Image analyzer and Quantity One software program were applied to quantify the total amount of ADAM-10. ADAM-10 protein was detected in both human and bovine cartilage. The strongest immunostaining was found in the cytoplasm and/or cell membranes of the superficial and upper middle layer of normal adult human cartilage, in the clusters and fibrillated areas of OA cartilage, and in IL-1alpha-stimulated bovine nasal cartilage. The distribution of ADAM-10 protein in bovine nasal cartilage was dependent on the length of exposure to IL-1alpha and corresponded to the areas of proteoglycan depletion. By Western blotting analysis of human cartilage, ADAM-10 was primarily detected in the membrane-enriched fraction and its levels were increased in degenerated and OA cartilage compared to normal cartilage. The results of this study suggest that ADAM-10 might be an important factor associated with cartilage degenerative processes. (J Histochem Cytochem 49:1165-1176, 2001)
Chubinskaya
S
S
Department of Biochemistry, Rush Medical College, Rush-Presbyterian-St Luke's Medical Center, 1653 W. Congress Parkway, Chicago, IL 60612, USA.
Mikhail
R
R
Deutsch
A
A
Tindal
M H
MH
D051722:ADAM Proteins; D000072197:ADAM10 Protein; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D053829:Amyloid Precursor Protein Secretases; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D015153:Blotting, Western; D002356:Cartilage / Q000378:metabolism* / Q000648:ultrastructure; D002358:Cartilage, Articular / Q000378:metabolism* / Q000648:ultrastructure; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000276:immunology / Q000378:metabolism*; D008875:Middle Aged; D009297:Nasal Mucosa / Q000378:metabolism*; D010003:Osteoarthritis / Q000378:metabolism*; D015854:Up-Regulation
D000911:Antibodies, Monoclonal; D007375:Interleukin-1; D008565:Membrane Proteins; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D008666:Metalloendopeptidases; C103778:ADAM 10 protein, Bos taurus; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human