Measurement of proteoglycan mRNA in articular cartilage--preliminary investigations.
Measurement of mRNA in adult articular cartilage could define whether treatment with nonsteroidal antiinflammatory drugs affects the rate of proteoglycan synthesis in vivo in osteoarthritis. We studied a variety of extraction and isolation methods to obtain a high yield of pure mRNA from adult articular cartilage. By combining a single step extraction procedure, precipitation in 0.8 M LiCl and poly d-T affinity isolation, we obtained pure mRNA that hybridized with a cDNA probe for the large aggregating proteoglycan. Thus, it should be feasible to assess the level of message using polymerase chain reaction technology; it is unlikely that northern blots, slot blots, or RNA protection assays can be performed directly on the small specimens of articular cartilage that are available from experimental model studies or from surgical specimens.
Adams
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ME
Department of Medicine, University of Calgary, AB, Canada.
Huang
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D000818:Animals; D002358:Cartilage, Articular / Q000737:chemistry* / Q000378:metabolism; D015342:DNA Probes; D004285:Dogs; D009693:Nucleic Acid Hybridization; D011509:Proteoglycans / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D012756:Sheep
D015342:DNA Probes; D011509:Proteoglycans; D012333:RNA, Messenger