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. 1974 Dec 1;63(3):843–854. doi: 10.1083/jcb.63.3.843

INFLUENCE OF EXTERNAL POTASSIUM ON THE SYNTHESIS AND DEPOSITION OF MATRIX COMPONENTS BY CHONDROCYTES IN VITRO

Jon C Daniel 1, Robert A Kosher 1, James E Hamos 1, James W Lash 1
PMCID: PMC2109377  PMID: 4279924

Abstract

The effect of a high external potassium concentration on the synthesis and deposition of matrix components by chondrocytes in cell culture was determined. There is a twofold increase in the amount of chondroitin 4- and 6-sulfate accumulated by chondrocytes grown in medium containing a high potassium concentration. There is also a comparable increase in the production of other sulfated glycosaminoglycans (GAG) including heparan sulfate and uncharacterized glycoprotein components. The twofold greater accumulation of GAG in the high potassium medium is primarily the result of a decrease in their rate of degradation. In spite of this increased accumulation of GAG, the cells in high potassium fail to elaborate appreciable quantities of visible matrix, although they do retain the typical chondrocytic polygonal morphology. Although most of the products are secreted into the culture medium in the high potassium environment, the cell layer retains the same amount of glycosaminoglycan as the control cultures. The inability of chondrocytes grown in high potassium to elaborate the typical hyaline cartilage matrix is not a consequence of an impairment in collagen synthesis, since there is no difference in the total amount of collagen synthesized by high potassium or control cultures. There is, however, a slight increase in the proportion of collagen that is secreted into the medium by chondrocytes in high potassium. Synthesis of the predominant cartilage matrix molecules is not sufficient in itself to ensure that these molecules will be assembled into a hyaline matrix.

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Selected References

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  1. Abbott J., Holtzer H. The loss of phenotypic traits by differentiated cells, V. The effect of 5-bromodeoxyuridine on cloned chondrocytes. Proc Natl Acad Sci U S A. 1968 Apr;59(4):1144–1151. doi: 10.1073/pnas.59.4.1144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ali S. Y., Evans L. Enzymic degradation of cartilage in osteoarthritis. Fed Proc. 1973 Apr;32(4):1494–1498. [PubMed] [Google Scholar]
  3. Coon H. G., Cahn R. D. Differentiation in vitro: effects of Sephadex fractions of chick embryo extract. Science. 1966 Sep 2;153(3740):1116–1119. doi: 10.1126/science.153.3740.1116. [DOI] [PubMed] [Google Scholar]
  4. Daniel J. C., Kosher R. A., Lash J. W., Hertz J. The synthesis of matrix components by chondrocytes in vitro in the presence of 5-bromodeoxyuridine. Cell Differ. 1973 Dec;2(5):285–298. doi: 10.1016/0045-6039(73)90033-x. [DOI] [PubMed] [Google Scholar]
  5. Douglas W. W. Stimulus-secretion coupling: the concept and clues from chromaffin and other cells. Br J Pharmacol. 1968 Nov;34(3):451–474. doi: 10.1111/j.1476-5381.1968.tb08474.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fratantoni J. C., Hall C. W., Neufeld E. F. The defect in Hurler's and Hunter's syndromes: faulty degradation of mucopolysaccharide. Proc Natl Acad Sci U S A. 1968 Jun;60(2):699–706. doi: 10.1073/pnas.60.2.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goggins J. F., Johnson G. S., Pastan I. The effect of dibutyryl cyclic adenosine monophosphate on synthesis of sulfated acid mucopolysaccharides by transformed fibroblasts. J Biol Chem. 1972 Sep 25;247(18):5759–5764. [PubMed] [Google Scholar]
  8. HAM R. G. CLONAL GROWTH OF MAMMALIAN CELLS IN A CHEMICALLY DEFINED, SYNTHETIC MEDIUM. Proc Natl Acad Sci U S A. 1965 Feb;53:288–293. doi: 10.1073/pnas.53.2.288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HOWELL D. S., DELCHAMPS E., RIEMER W., KIEM I. A profile of electrolytes in the cartilaginous plate of growing ribs. J Clin Invest. 1960 Jun;39:919–929. doi: 10.1172/JCI104112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hascall V. C., Sajdera S. W. Proteinpolysaccharide complex from bovine nasal cartilage. The function of glycoprotein in the formation of aggregates. J Biol Chem. 1969 May 10;244(9):2384–2396. [PubMed] [Google Scholar]
  11. Hinegardner R. T. An improved fluorometric assay for DNA. Anal Biochem. 1971 Jan;39(1):197–201. doi: 10.1016/0003-2697(71)90476-3. [DOI] [PubMed] [Google Scholar]
  12. Holthausen H. S., Chacko S., Davidson E. A., Holtzer H. Effect of 5-bromodeoxyuridine on expression of cultured chondrocytes grown in vitro. Proc Natl Acad Sci U S A. 1969 Jul;63(3):864–870. doi: 10.1073/pnas.63.3.864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. KISSANE J. M., ROBINS E. The fluorometric measurement of deoxyribonucleic acid in animal tissues with special reference to the central nervous system. J Biol Chem. 1958 Jul;233(1):184–188. [PubMed] [Google Scholar]
  14. Kojima K., Yamagata T. Glycosaminoglycans and electrokinetic behavior of rat ascites hepatoma cells. Exp Cell Res. 1971 Jul;67(1):142–146. doi: 10.1016/0014-4827(71)90629-x. [DOI] [PubMed] [Google Scholar]
  15. Kosher R. A., Searls R. L. Sulfated mucopolysaccharide synthesis during the development of Rana pipiens. Dev Biol. 1973 May;32(1):50–68. doi: 10.1016/0012-1606(73)90219-4. [DOI] [PubMed] [Google Scholar]
  16. Kraemer P. M. Heparan sulfates of cultured cells. I. Membrane-associated and cell-sap species in Chinese hamster cells. Biochemistry. 1971 Apr 13;10(8):1437–1445. doi: 10.1021/bi00784a026. [DOI] [PubMed] [Google Scholar]
  17. Kraemer P. M. Heparan sulfates of cultured cells. II. Acid-soluble and -precipitable species of different cell lines. Biochemistry. 1971 Apr 13;10(8):1445–1451. doi: 10.1021/bi00784a027. [DOI] [PubMed] [Google Scholar]
  18. LAGUNOFF D., WARREN G. Determination of 2-deoxy-2-sulfoaminohexose content of mucopolysaccharides. Arch Biochem Biophys. 1962 Dec;99:396–400. doi: 10.1016/0003-9861(62)90285-0. [DOI] [PubMed] [Google Scholar]
  19. Lash J. W., Rosene K., Minor R. R., Daniel J. C., Kosher R. A. Environmental enhancement of in vitro chondrogenesis. 3. The influence of external potassium ions and chondrogenic differentiation. Dev Biol. 1973 Dec;35(2):370–375. doi: 10.1016/0012-1606(73)90032-8. [DOI] [PubMed] [Google Scholar]
  20. Matalon R., Dorfman A. Hurler's syndrome, an -L-iduronidase deficiency. Biochem Biophys Res Commun. 1972 May 26;47(4):959–964. doi: 10.1016/0006-291x(72)90586-4. [DOI] [PubMed] [Google Scholar]
  21. Miller E. J., Martin G. R., Piez K. A., Powers M. J. Characterization of chick bone collagen and compositional changes associated with maturation. J Biol Chem. 1967 Dec 10;242(23):5481–5489. [PubMed] [Google Scholar]
  22. Miller E. J., Matukas V. J. Chick cartilage collagen: a new type of alpha 1 chain not present in bone or skin of the species. Proc Natl Acad Sci U S A. 1969 Dec;64(4):1264–1268. doi: 10.1073/pnas.64.4.1264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Minor R. R. Somite chondrogenesis. A structural analysis. J Cell Biol. 1973 Jan;56(1):27–50. doi: 10.1083/jcb.56.1.27. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Neufeld E. F., Cantz M. J. Corrective factors for inborn errors of mucopolysaccharide metabolism. Ann N Y Acad Sci. 1971 Jul 6;179:580–587. doi: 10.1111/j.1749-6632.1971.tb46934.x. [DOI] [PubMed] [Google Scholar]
  25. Peterkofsky B., Diegelmann R. Use of a mixture of proteinase-free collagenases for the specific assay of radioactive collagen in the presence of other proteins. Biochemistry. 1971 Mar 16;10(6):988–994. doi: 10.1021/bi00782a009. [DOI] [PubMed] [Google Scholar]
  26. Peterkofsky B. The effect of ascorbic acid on collagen polypeptide synthesis and proline hydroxylation during the growth of cultured fibroblasts. Arch Biochem Biophys. 1972 Sep;152(1):318–328. doi: 10.1016/0003-9861(72)90221-4. [DOI] [PubMed] [Google Scholar]
  27. SANTOIANNI P., AYALA M. FLUOROMETRIC ULTRAMICROANALYSIS OF DEOXYRIBONUCLEIC ACID IN HUMAN SKIN. J Invest Dermatol. 1965 Aug;45:99–103. doi: 10.1038/jid.1965.100. [DOI] [PubMed] [Google Scholar]
  28. Saito H., Yamagata T., Suzuki S. Enzymatic methods for the determination of small quantities of isomeric chondroitin sulfates. J Biol Chem. 1968 Apr 10;243(7):1536–1542. [PubMed] [Google Scholar]
  29. Shulman H. J., Meyer K. Protein-polyaccharide of chicken cartilage and chondrocyte cell cultures. Biochem J. 1970 Dec;120(4):689–697. doi: 10.1042/bj1200689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Slomiany B. L., Meyer K. Isolation and structural studies of sulfated glycoproteins of hog gastric mucosa. J Biol Chem. 1972 Aug 25;247(16):5062–5070. [PubMed] [Google Scholar]
  31. Suzuki S., Kojima K., Utsami K. R. Production of sulfated mucopolysaccharides by established cell lines of fibroblastic and nonfibroblastic origin. Biochim Biophys Acta. 1970 Oct 27;222(1):240–243. doi: 10.1016/0304-4165(70)90377-6. [DOI] [PubMed] [Google Scholar]
  32. Toole B. P., Gross J. The extracellular matrix of the regenerating newt limb: synthesis and removal of hyaluronate prior to differentiation. Dev Biol. 1971 May;25(1):57–77. doi: 10.1016/0012-1606(71)90019-4. [DOI] [PubMed] [Google Scholar]
  33. Woessner J. F., Jr Cartilage cathepsin D and its action on matrix components. Fed Proc. 1973 Apr;32(4):1485–1488. [PubMed] [Google Scholar]
  34. Woessner J. F., Jr Cartilage cathepsin D and its action on matrix components. Fed Proc. 1973 Apr;32(4):1485–1488. [PubMed] [Google Scholar]

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