Effect of Ethanol on Neural Cells Grown in Culture: Interaction with Plasma Membrane Ecto-5’-Nucleotidase Activity

  • Peter J. Syapin
  • Sujata Tewari
  • Ernest P. Noble
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 126)


The effect of ethanol on an enzyme system within an intact functional plasma membrane has been studied using neural cells grown in culture. Rat C6 glioma cells in monolayer culture were treated acutely or chronically with 100 mM ethanol and the effect of this exposure on the activity of ecto-5’-nucleotidase was determined. Acute exposure led to an increase in enzyme activity with maximum stimulation occurring at concentrations of 100 — 400 mM ethanol. Chronic treatment of cells with 100 mM ethanol for 4 — 8 days also caused an increase in ecto-5’-nucleotidase activity. Both the acute and chronic ethanol-induced stimulation of enzyme activity was completely reversible by removing the ethanol; the acute effects reversed immediately, whereas the chronic effects required several hours. The addition of Concanavalin A demonstrated that the effects on enzyme activity of both chronic and acute exposure to ethanol were blocked by modification of the external cell surface. The effect of chronic exposure to 100 mM ethanol was further localized to an action on the plasma membrane by studies which showed chronic exposure to have no effect on the intracellular 5’-nucleotidase activity. Furthermore, the occurrence of pharmacological tolerance to acute ethanol was observed in this plasma membrane system following chronic treatment of C6 cells with 100 mM ethanol. These findings are consistent with the hypothesis that mammalian neural cells can adapt to the chronic presence of ethanol through changes in their plasma membrane.


Neural Cell Ethanol Exposure Chronic Ethanol Cell Plasma Membrane Acute Ethanol 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Benda, P., Lightbody, L., Sato, G., Levine, L., and Sweet, W. Differentiated rat glial cell strains in tissue culture. Science 161 370–371, 1968.PubMedCrossRefGoogle Scholar
  2. Chin, J.H., and Goldstein, D.B. Effects of low concentrations of ethanol on the fluidity of spin labeled erythrocyte and brain membranes. Mol. Pharmacol., 13:435–441, 1977a.PubMedGoogle Scholar
  3. Chin, J.H., and Goldstein, D.B. Drug tolerance in biomembranes: A spin label study of the effects of ethanol. Science, 196:684–685, 1977b.PubMedCrossRefGoogle Scholar
  4. Curran, M., and Seeman, P. Alcohol tolerance in a cholinergic nerve terminal: Relation to the membrane expansion-fluidization theory of alcohol action. Science, 197:910–911, 1977.PubMedCrossRefGoogle Scholar
  5. De Pierre, J.W., and Karnovsky, M.L. Plasma membrane of mammalian cells: A review of methods for their characterization and isolation. J. Cell Biol., 56:275–303, 1973.CrossRefGoogle Scholar
  6. DePierre, J.W., and Karnovsky, M.L. Ecto-enzymes of the guinea pig polymorphonuclear leukocyte: I. Evidence for an ecto-adenosine monophosphatase, adenosine triphosphatase and p-nitrophenyl phosphate. J. Biol. Chem., 56:7111–7120, 1977.Google Scholar
  7. Gomori, G. A modification of the colorimetric phosphorous determination for use with the photoelectric colorimeter. J. Lab. Clin. Med., 27:955–960, 1942.Google Scholar
  8. Gurd, J.W., and Evans, H. Distribution of liver plasma membrane 5’-nucleotidase as indicated by its reaction with anti-plasma membrane serum. Arch. Biochem. Biophys., 164:305–311, 1974.PubMedCrossRefGoogle Scholar
  9. Hill, M.W., and Bangham, A.B. General depressant drug dependency: A biophysical hypothesis. In M.M. Gross (ed.) Alcohol Intoxication and Withdrawal II, pp. 1–9, Plenum Press, New York, 1975.Google Scholar
  10. Hunt, W.A. The effects of aliphatic alcohols on the biophysical and biochemical correlates of membrane function. In E. Majchrowicz (ed.) Biochemical Pharmacology of Ethanol, pp. 195–212, Plenum Press, New York, 1975.CrossRefGoogle Scholar
  11. Kalant, H. Absorption, diffusion, distribution and elimination of ethanol: Effects on biological membranes. In B. Kissin and H. Begleiter (eds.) The Biology of Alcoholism, Vol. 1, pp. 1–62, Plenum Press, New York, 1971.CrossRefGoogle Scholar
  12. Kalant, H., and Grose, W. Effects of ethanol and pentobarbital on release of acetylcholine from cerebral cortex slices. J. Pharmacol. Exp. Ther., 158:386–393, 1967.PubMedGoogle Scholar
  13. Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J. Protein measurement with the Folin phenol reagent. J. Biol. Chem., 193; 265–275, 1951.PubMedGoogle Scholar
  14. Massarelli, R., Syapin, P.J., and Noble, E.P. Increased uptake of choline by neural cell cultures chronically exposed to ethanol. Life Sei., 18:397–404, 1976.CrossRefGoogle Scholar
  15. Newby, A.C., Luzio, J.P., and Hales, C.N. The properties and extracellular location of 5’-nucleotidase of the rat fat cell plasma membrane. Biochem. J., 146:625–633, 1975.PubMedGoogle Scholar
  16. Noble, E.P., Syapin, P.J., Vigran, R., and Rosenberg, A., Neuroaminidase-releasable surface sialic acid of cultured astroblasts exposed to ethanol. J. Neurochem., 27:217–221, 1976.PubMedCrossRefGoogle Scholar
  17. Noble, E.P., and Tewari, S. Metabolic aspects of alcohol in the brain. In. C.S. Lieber (ed.) Metabolic Aspects of Alcoholism, pp. 149–185, University Park Press, Baltimore, 1976.Google Scholar
  18. So, L.L., and Goldstein, I.J. Protein-carbohydrate interaction. XX. On the number of combining sites on concanavalin A, the phytohemagglutinin of the Biochem. Biophys. Acta, 165:398–414, 1968.Google Scholar
  19. Stefanovic, V., Mandel, P., and Rosenberg, A. Concanavalin A inhibition of ecto-5’-nucleotidase of intact cultured C6 glioma cells. J. Biol. Chem., 250:7081–7083, 1975.PubMedGoogle Scholar
  20. Stefanovic, V., Mandel, P., and Rosenberg, A. Ecto-5’-nucleotidase of intact culturec C6 rat glioma cells. J. Biol. Chem., 251: 3900–3905, 1976.PubMedGoogle Scholar
  21. Syapin, P.J. Studies on acute and chronic ethanol treatment and the plasma membrane activity of neural cells in monolayer culture. Ph.D. Dissertation, University of California, Irvine, 1975.Google Scholar
  22. Syapin, P.J., and Noble, E.P. Studies of ethanol effects on cells in culture. In E. Majchrowicz and E.P. Noble (eds.) Biochemical Pharmacology of Ethanol II, Plenum Press, New York, 1978 (in press).Google Scholar
  23. Syapin, P.J., Stefanovic, V., Ciesielski-Treska, J., Mandel, P., and Noble, E.P. The effects of ethanol on growth and ectoenzyme activity of cultured cells (Abst.), Fifth International Society for Neurochemistry, p. 232, Barcelona, Spain, 1975.Google Scholar
  24. Syapin, P.J., Stefanovic, V., Mandel, P., and Noble, E.P. The chronic and acute effects of ethanol on adenosine triphosphatase activity in cultured astroblast and neuroblastoma cells. J. Neurosci. Res., 2:147–155, 1976.PubMedCrossRefGoogle Scholar
  25. Trams, E.G., and Lauter, C.J. On the sidedness of plasma membrane enzymes. Biochim. Biophys. Acta, 345: 180–197, 1974.PubMedCrossRefGoogle Scholar
  26. Traynor, M.E., Woodson, P.B.J., Schlapfer, W.T., and Barondes, S.H. Sustained tolerance to a specific effect of ethanol on post- tetanic potentiation in Aphysia, Science, 193:510–511, 1976.PubMedCrossRefGoogle Scholar
  27. Wallgren, H. Effects of ethanol on intracellular respiration and cerebral function. In B. Kissin and H. Begleiter (eds.) The Biology of Alcoholism, Vol. 1, pp. 103–125, Plenum Press, New York, 1971.CrossRefGoogle Scholar
  28. Wallgren, H., and Barry, H. Actions of Alcohol, Vol. 1, p. 56, Elsevier, Amsterdam, 1970.Google Scholar

Copyright information

© Plenum Press, New York 1980

Authors and Affiliations

  • Peter J. Syapin
    • 1
    • 2
  • Sujata Tewari
    • 1
    • 2
  • Ernest P. Noble
    • 1
    • 2
  1. 1.Neurochemistry Laboratories Department of Psychiatry and Human BehaviorUniversity of CaliforniaIrvineUSA
  2. 2.National Institute on Alcohol Abuse and AlcoholismRockvilleUSA

Personalised recommendations