Abstract
Several methods have been described for investigating chondrocyte metabolism in vitro. In this study, in-situ hybridization (ISH) using an oligonucleotide probe (i.e. a poly-d(T) probe) to detect total messenger RNA (mRNA) in cartilage explants has been compared with radiosulphate and radioleucine uptake studies in an attempt to assess the value of ISH in investigating chondrocyte metabolism. The relative results of the three parameters indicate qualitative similarities in cells in the intermediate, deep and calcified zones but differences in the superficial zone. The relative levels of mRNA and leucine and sulphate uptake in the midzone areas could be construed as indicating that the bulk of cellular activity was directed towards the synthesis of proteoglycans. A similar relation between the three parameters, but at a lower level, was seen in chondrocytes in the calcified zone demonstrating that these cells are viable and biosynthetic. Both quantitative and qualitative differences between the three methods were observed in the superficial chondrocytes regarding the amount of mRNA compared to sulphate and leucine uptake. The results suggest that ISH can detect differences in the amount of mRNA present in chondrocytes in differing zones of cartilage and, like the radioleucine and radiosulphate studies, particularly emphasizes their functional heterogeneity.
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- Bayliss M. T., Venn M., Maroudas A., Ali S. Y. Structure of proteoglycans from different layers of human articular cartilage. Biochem J. 1983 Feb 1;209(2):387–400. doi: 10.1042/bj2090387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Björnsson S., Heinegård D. Fractionation and characterization of proteoglycans isolated from chondrocyte cell cultures. Biochem J. 1981 Aug 1;197(2):249–258. doi: 10.1042/bj1970249. [DOI] [PMC free article] [PubMed] [Google Scholar]
- CURRAN R. C., GIBSON T. The uptake of labelled sulphate by human cartilage cells and its use as a test for viability. Proc R Soc Lond B Biol Sci. 1956 Mar 13;144(917):572–576. doi: 10.1098/rspb.1956.0012. [DOI] [PubMed] [Google Scholar]
- Dunham J., Shackleton D. R., Nahir A. M., Billingham M. E., Bitensky L., Chayen J., Muir I. H. Altered orientation of glycosaminoglycans and cellular changes in the tibial cartilage in the first two weeks of experimental canine osteoarthritis. J Orthop Res. 1985;3(3):258–268. doi: 10.1002/jor.1100030302. [DOI] [PubMed] [Google Scholar]
- Hascall V. C., Handley C. J., McQuillan D. J., Hascall G. K., Robinson H. C., Lowther D. A. The effect of serum on biosynthesis of proteoglycans by bovine articular cartilage in culture. Arch Biochem Biophys. 1983 Jul 1;224(1):206–223. doi: 10.1016/0003-9861(83)90205-9. [DOI] [PubMed] [Google Scholar]
- Jilbert A. R., Burrell C. J., Gowans E. J., Rowland R. Histological aspects of in situ hybridization. Detection of poly(A) nucleotide sequences in mouse liver sections as a model system. Histochemistry. 1986;85(6):505–514. doi: 10.1007/BF00508433. [DOI] [PubMed] [Google Scholar]
- Lamb R. A., Choppin P. W. The gene structure and replication of influenza virus. Annu Rev Biochem. 1983;52:467–506. doi: 10.1146/annurev.bi.52.070183.002343. [DOI] [PubMed] [Google Scholar]
- Lewis M. E., Sherman T. G., Watson S. J. In situ hybridization histochemistry with synthetic oligonucleotides: strategies and methods. Peptides. 1985;6 (Suppl 2):75–87. doi: 10.1016/0196-9781(85)90138-x. [DOI] [PubMed] [Google Scholar]
- Parkkinen J. J., Paukkonen K., Pesonen E., Lammi M. J., Markkanen S., Helminen H. J., Tammi M. Quantitation of autoradiographic grains in different zones of articular cartilage with image analyzer. Histochemistry. 1990;93(3):241–245. doi: 10.1007/BF00266384. [DOI] [PubMed] [Google Scholar]
- Warford A. In situ hybridisation: a new tool in pathology. Med Lab Sci. 1988 Oct;45(4):381–394. [PubMed] [Google Scholar]