Expression of hyaluronan synthases in articular cartilage.
OBJECTIVE
To investigate the mRNA expression profiles of three mammalian hyaluronan synthases (HAS1, HAS2 and HAS3) in chondrocytes from normal (undiseased) animal cartilage and osteoarthritic human cartilage maintained in experimental culture systems and exposed to catabolic or anabolic stimuli provided by cytokines, growth factors and retinoic acid.


METHODS
Chondrocytes isolated from normal bovine, porcine or from osteoarthritic human cartilage were cultured as monolayers or embedded in agarose. Cultures were maintained for 3-5 days in the presence or absence of catabolic stimuli (IL-1, TNF-alpha or retinoic acid) or anabolic stimuli (TGF-beta or IGF-1) followed by extraction of RNA and analysis of HAS mRNA expression by RT-PCR.


RESULTS
Whereas mRNA for HAS1 was not detected in any sample, the mRNAs for HAS2 and HAS3 were expressed in human, bovine and porcine chondrocytes. HAS2 mRNA was present in chondrocytes from all cartilages and under all culture conditions, whereas HAS3 did not show such constitutive expression. In agarose cultures of bovine and porcine chondrocytes HAS2 mRNA was present in control, IL-1 and retinoic acid treated cultures, whereas HAS3 mRNA was only detected in IL-1 stimulated cultures. Mature bovine chondrocytes cultured in monolayers expressed mRNAs for both HAS2 and HAS3 in the presence of IL-1, TNF-alpha, TGF-beta and IGF-1, however immature bovine chondrocytes in monolayer cultures displayed virtually no HAS3 mRNA expression in the presence of these cytokines and growth factors. HAS2 and HAS3 mRNAs were also expressed by bovine chondrocytes isolated from either the superficial or deep zone of articular cartilage, and by human chondrocytes cultured either in the absence or presence of IL-1 and retinoic acid.


CONCLUSIONS
Our data indicate that HAS2 and HAS3 (but not HAS1) mRNAs are expressed in several mammalian cartilages. Chondrocyte HAS2 mRNA appears to be constitutively expressed while chondrocyte HAS3 mRNA expression can be differentially regulated in an age-dependent fashion, and may be affected by local and/or systemic catabolic or anabolic stimuli provided by cytokines or growth factors.
Hiscock
D R
DR
Connective Tissue Biology Laboratories, Cardiff School of Biosciences, Cardiff University, Wales, UK.
Caterson
B
B
Flannery
C R
CR
D000368:Aged; D000375:Aging / Q000378:metabolism; D000818:Animals; D002358:Cartilage, Articular / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000235:genetics*; D006801:Humans; D000076002:Hyaluronan Synthases; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D008322:Mammals / Q000378:metabolism*; D020370:Osteoarthritis, Knee / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013552:Swine; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D007375:Interleukin-1; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D014212:Tretinoin; D007334:Insulin-Like Growth Factor I; D014453:Glucuronosyltransferase; C101754:HAS2 protein, human; C105458:HAS3 protein, human; D000076002:Hyaluronan Synthases