Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1968 Apr 1;37(1):13–26. doi: 10.1083/jcb.37.1.13

LOCALIZATION OF HYALURONIC ACID IN SYNOVIAL CELLS BY RADIOAUTOGRAPHY

Peter Barland 1, Carol Smith 1, David Hamerman 1
PMCID: PMC2107396  PMID: 5689533

Abstract

Cultured human synovial cells secrete hyaluronic acid (HA) into the culture medium. Glucosamine-6-3H was shown to be a direct and relatively specific precursor of HA-3H by the following observations: the susceptibility of nondialyzable radioactivity in the medium to hyaluronidase, its migration with hexuronic acid on zone electrophoresis in polyvinyl chloride, its exclusion from Sephadex G-200, and the localization of radioactivity to glucosamine after hydrolysis of the labeled polysaccharide. The presence of intracellular HA-3H was established by sequential extraction of labeled cells and by radioautography of synovial cell cultures digested with hyaluronidase in situ. When cells were exposed to medium lacking glucose, glucosamine-3H-uptake was enhanced; and this made possible electron microscopic radioautographic studies. These studies demonstrate the early and continued presence of HA-3H within the Golgi apparatus.

Full Text

The Full Text of this article is available as a PDF (1.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BITTER T., MUIR H. M. A modified uronic acid carbazole reaction. Anal Biochem. 1962 Oct;4:330–334. doi: 10.1016/0003-2697(62)90095-7. [DOI] [PubMed] [Google Scholar]
  2. CARO L. G., PALADE G. E. PROTEIN SYNTHESIS, STORAGE, AND DISCHARGE IN THE PANCREATIC EXOCRINE CELL. AN AUTORADIOGRAPHIC STUDY. J Cell Biol. 1964 Mar;20:473–495. doi: 10.1083/jcb.20.3.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. CASTOR C. W., FRIES F. F. Composition and function of human synovial connective tissue cells measured in vitro. J Lab Clin Med. 1961 Mar;57:394–407. [PubMed] [Google Scholar]
  4. CRANE R. K., FIELD R. A., CORI C. F. Studies of tissue permeability. I. The penetration of sugars into the Ehrlich ascites tumor cells. J Biol Chem. 1957 Feb;224(2):649–662. [PubMed] [Google Scholar]
  5. GODMAN G. C., LANE N. ON THE SITE OF SULFATION IN THE CHONDROCYTE. J Cell Biol. 1964 Jun;21:353–366. doi: 10.1083/jcb.21.3.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GROSSFELD H., MEYER K., GODMAN G., LINKER A. Mucopolysaccharides produced in tissue culture. J Biophys Biochem Cytol. 1957 May 25;3(3):391–396. doi: 10.1083/jcb.3.3.391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hamerman D., Todaro G. J., Green H. The production of hyaluronate by spontaneously established cell lines and viral transformed lines of fibroblastic origin. Biochim Biophys Acta. 1965 Nov 1;101(3):343–351. doi: 10.1016/0926-6534(65)90013-8. [DOI] [PubMed] [Google Scholar]
  8. Kornfeld S., Ginsburg V. The metabolism of glucosamine by tissue culture cells. Exp Cell Res. 1966 Mar;41(3):592–600. doi: 10.1016/s0014-4827(66)80109-x. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Lawford G. R., Schachter H. Biosynthesis of glycoprotein by liver. The incorporation in vivo of 14C-glucosamine into protein-bound hexosamine and sialic acid of rat liver subcellular fractions. J Biol Chem. 1966 Nov 25;241(22):5408–5418. [PubMed] [Google Scholar]
  11. MORRIS C. C. Quantitative studies on the production of acid mucopolysaccharides by replicate cell cultures of rat fibroblasts. Ann N Y Acad Sci. 1960 Jun 30;86:878–915. doi: 10.1111/j.1749-6632.1960.tb42848.x. [DOI] [PubMed] [Google Scholar]
  12. Molnar J., Lutes R. A., Winzler R. J. The biosynthesis of glycoproteins. V. Incorporation of glucosamine-1-14 C into macromolecules by Ehrlich ascites carcinoma cells. Cancer Res. 1965 Oct;25(9):1438–1445. [PubMed] [Google Scholar]
  13. Neutra M., Leblond C. P. Radioautographic comparison of the uptake of galactose-H and glucose-H3 in the golgi region of various cells secreting glycoproteins or mucopolysaccharides. J Cell Biol. 1966 Jul;30(1):137–150. doi: 10.1083/jcb.30.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Peters T., Jr, Ashley C. A. An artefact in radioautography due to binding of free amino acids to tissues by fixatives. J Cell Biol. 1967 Apr;33(1):53–60. doi: 10.1083/jcb.33.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. ROBINSON G. B., MOLNAR J., WINZLER R. J. BIOSYNTHESIS OF GLYCOPROTEINS. I. INCORPORATION OF GLUCOSAMINE-14C INTO LIVER AND PLASMA PROTEINS OF THE RAT. J Biol Chem. 1964 Apr;239:1134–1141. [PubMed] [Google Scholar]
  16. SALPETER M. M., BACHMANN L. AUTORADIOGRAPHY WITH THE ELECTRON MICROSCOPE. A PROCEDURE FOR IMPROVING RESOLUTION, SENSITIVITY, AND CONTRAST. J Cell Biol. 1964 Aug;22:469–477. doi: 10.1083/jcb.22.2.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. SARCIONE E. J., BOHNE M., LEAHY M. THE SUBECELLULAR SITE OF HEXOSAMINE INCORPORATION INTO LIVER PROTEIN. Biochemistry. 1964 Dec;3:1973–1976. doi: 10.1021/bi00900a032. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES