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Glycolipids in Cultured Fetal Tay-Sachs Disease Cerebellar Cells

  • Larry Schneck
  • Linda M. Hoffman
  • Daniel Amsterdam
  • Steven Brooks
  • Betty Pinkett
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 68)

Abstract

Tay-Sachs disease (TSD) is a fatal, genetically determined disorder of sphingoglycolipid metabolism, associated with the absence of the lysosomal enzyme B-D-N-acetylhexosaminidase A (Hex A) (1). There is a massive accumulation of GM2 ganglioside, and its asialo derivative GA2 in cells of the central nervous system. A cell culture which reproduces these biochemical parameters would permit one to measure the effect of enzyme replacement therapy under controlled conditions that are not easily attainable in vivo. Although skin fibroblasts cultured from TSD patients lack Hex A, these cultured cells do not accumulate GM2 ganglioside (2). Since TSD is a neuronal lipid storage disease, and since fetal TSD brain has the characteristic glycolipid patterns found in infant TSD brain, a cell strain from fetal TSD cerebellum was established and the glycolipid patterns were evaluated by TLC and GLC (3). The cells were labelled with 14C-glucosamine in order to compare the metabolic activity of the gangliosides over an extended time period. The cells were also transformed with the oncogenic DNA SV-40 virus (4) since we wished to establish a permanent cell line for the study of this disease.

Keywords

Enzyme Replacement Therapy Fetal Brain Hydroxy Fatty Acid Infant Brain Cerebellar Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Okada, S., and O’Brien, J. S., Tay-Sachs disease: Generalized absence of a Beta-D-N-Acetylhexosaminidase component, Science 165 (1969) 698–700.PubMedCrossRefGoogle Scholar
  2. 2.
    Dawson, G., Matalon, R., and Dorfman, A., Glycosphingolipids in cultured human fibroblasts. II. Characterization and metabolism in fibroblasts from patients with errors of glycosphingolipid and mucopolysaccharide metabolism. J. Biol. Chem., 247 (1972) 5951–5958.PubMedGoogle Scholar
  3. 3.
    Hoffman, L. M., Amsterdam, D., and Schneck, L., GM2 ganglioside in fetal Tay-Sachs disease brain cell culture: A model system for the disease. Brain Res. (1975) In press.Google Scholar
  4. 4.
    Shein, H. M., Transformation of astrocytes and destruction of spongioblasts induced by Simian tumor virus (SV40) in cultures of human fetal neuroglia, J. Neuropath. & Exp. Neurol. 26 (1967) 60–76.CrossRefGoogle Scholar
  5. 5.
    Amsterdam, D., and Brooks, S. E., Methodology: Cell culture. In B. W. Volk and L. Schneck (Eds.), The Gangliosidoses, Plenum Press, New York, 1975, pp. 265–270.Google Scholar
  6. 6.
    Suzuki, K., The pattern of mammalian brain gangliosides. II. Evaluation of the extraction procedures, post-mortem changes and the effect of formalin preservation, J. Neurochem. 12 (1965) 629–638.PubMedCrossRefGoogle Scholar
  7. 7.
    Folch, J., Lees, M., and Sloane-Stanley, G. H., A simple method for the isolation and purification of total lipides from animal tissues, J. Biol. Chem., 226 (1957) 497–509.PubMedGoogle Scholar
  8. 8.
    Schneck, L., Pihkett, B., and Volk, B. W., Asialo GM2-gan-glioside in fetal Tay-Sachs disease brain, J. Neurochem. 24 (1975) 183–184.PubMedCrossRefGoogle Scholar
  9. 9.
    Sandhoff, K., The hydrolysis of Tay-Sachs ganglioside (TSG) by human N-acetyl-B-D-hexosaminidase A, FEBS Lett. 11 (1970) 342–344.PubMedCrossRefGoogle Scholar
  10. 10.
    Wenger, D. A., Okada, S., and O’Brien, J. S., Studies on the substrate specificity of hexosaminidase A and B from liver, Arch. Biochem. Biophys. 153 (1972) 116–129.PubMedCrossRefGoogle Scholar
  11. 11.
    Greenbaum, M., Hoffman, L. M., and Schneck, L., Unpublished observations.Google Scholar
  12. 12.
    Sandhoff, K., Harzer, K., Wassle, W., and Jatzkewitz, H., Enzyme alterations and lipid storage in three variants of Tay-Sachs disease, J. Neurochem. 18 (1971) 2469–2489.PubMedCrossRefGoogle Scholar
  13. 13.
    Navon, R., Padeh, B. and Adam, A., Apparent deficiency of hexosaminidase A in healthy members of a family with Tay-Sachs disease, Am. J. Hum. Genet. 25 (1973) 287–293.PubMedGoogle Scholar
  14. 14.
    Yanier, M. T., Holm, M., Mansson, J. E., and Svennerholm, L., Gangliosides of infant brain, J. Neurochem. 21 (1973) 1375–1384.CrossRefGoogle Scholar
  15. 15.
    Robert, J., Freysz, L., Sensenbrenner, M., Mandel, P., and Rebel, G., Gangliosides of glial cells: A comparative study of normal astroblasts in tissue culture and glial cells isolated on sucrose-ficoll gradients, FEBS Lett. 50 (1975) 144–146.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1976

Authors and Affiliations

  • Larry Schneck
    • 1
  • Linda M. Hoffman
    • 1
  • Daniel Amsterdam
    • 1
  • Steven Brooks
    • 1
  • Betty Pinkett
    • 1
  1. 1.Department of Neurology and the Isaac Albert ResearchInstitute of the Kingsbrook Jewish Medical CenterBrooklynUSA

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