Organized Cultures of Nerve Tissue: A Novel Model System for Studies of Lipid Protein Interaction on the Functional Level

  • Michael Giesing
Part of the NATO Advanced Science Institutes Series book series (NSSA, volume 71)


Organized fragments of grey matter of cerebral cortex (CC) maintained in vitro during cell maturation in the explant culture assembly are presented as a novel tool for studies on the functional impact of membrane phospholipids (PL). PL have been found to affect two transport systems and one receptor system. The specificity of the PL effects has been examined by analyzing the kinetics of proteins that bind glycine — an irrelevant amino acid neurotransmitter in CC — and γ-aminobutyric acid (GABA), a major inhibitory neurotransmitter in the tissue. The composition of membrane constituents has been changed either through introduction of exogenous PL into the lipid matrix of viable cells or through specific degradation of asymmetrically distributed components. The following findings that were made are discussed:
  1. 1)

    The glycine carrier is a transversal protein that is governed by phosphatidylcholine (PC) but not by phosphatidylethanolamine (PE) in a fatty acid specific fashion. PC binds the ligand as well as it affects the protein. The carrier does not undergo lateral motion between ordered and fluid lipid domains. On the whole the glycine carrier is a protein with relatively little functional relation to the compositional mosaicism of the plasma membrane.

  2. 2)

    The GABA carrier is a mobile protein that seems to be localized to a major extent in the outer leaflet. Asymmetrically distributed PL regulate the activity of the protein in a fatty acid specific manner. The carrier can be inhibited as well as stimulated.

  3. 3)

    The GABA receptor is not a lipoprotein in nature. PL affect the receptor activity either through a single transversal modulator protein or through two asymmetrically distributed modulator proteins. PL that are components of the outer leaflet such as phosphatidyl-N-dimethylethanolamine (PDE) activate the inhibitory capacity of the modulator leading to a state of desensitization of the receptor that is characterized by an increase in the strength of ligand binding. Cytoplasmic PL, among them mainly PE and phosphatidyl-N-monoethanolamine (PME), induce a stimulation of GABA receptor binding through activation of the inner part of the transversal modulator or of the cytoplasmic modulator, thus creating a state of super-sensitivity. Cholinergic excess stimulation of the network of cultured neurons by bath application of carbachol increases PDE formation via the methylation pathway. This is accompanied by desensitization of the GABA receptor. The state of desensitization should be terminated through PE. The findings illustrate the functional specificity of PL in nerve tissue. It seems to be essential that viable cells are used since results stemming from artificial membranes of membrane preparations are not always the same.



Gaba Receptor Polar Head Outer Leaflet Viable Nerve Cell Gaba Transport 
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.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    M.B. Anand-Srivastava and R.A. Johnson, Role of phospholipids in coupling of adenosine and dopamine receptors to striatal adenylate cyclase, J. Neurochem. 36: 1819 (1981).PubMedCrossRefGoogle Scholar
  2. 2.
    T.J. Andreasen, D.R. Doerge, and M.G. McNamee, Effect of phospholipase A2 on the binding and ion permeability control properties of the acetylcholine receptor, Arch. Biochem. Biophys. 194: 468 (1979).PubMedCrossRefGoogle Scholar
  3. 3.
    A.H. Aprison and E.C. Daly, Biochemical aspects of transmission at inhibitory synapses: The role of glycine, in: “Advances in Neurochemistry”, Vol.3, B.W. Agranoff and M.H. Aprison, eds., Plenum Press, New York, pp. 203 (1978).CrossRefGoogle Scholar
  4. 4.
    V.J.Balcar, J. Berg, J. Robert, and P. Mandel, Uptake of L-glutamate and taurine in neuroblastoma cells with altered fatty acid composition of membrane phospholipids, J. Neurochem. 34: 1678 (1980).PubMedCrossRefGoogle Scholar
  5. 5.
    V.J.Balcar and P. Mandel, Inhibition of high affinity uptake of GABA by branched fatty acids, Experientia, 32: 904 (1976).PubMedCrossRefGoogle Scholar
  6. 6.
    N.G. Bazan, Effects of ischemia and electroconvulsive shock on free fatty acid pool in the brain, Biochim. Biophys. Acta, 218: 1 (1970).PubMedGoogle Scholar
  7. 7.
    T.H. Chiu and H.C. Rosenberg, Endogenous modulator of benzodiazepine binding in rat cortex, J. Neurochem. 36: 336 (1981).PubMedCrossRefGoogle Scholar
  8. 8.
    A.Y. Chwelch and S.W. Uslie, Phosphatidylserine enhancement of [3H] γ-aminobutyric acid uptake by rat whole brain synaptosomes, J. Neurochem. 38: 691 (1982).CrossRefGoogle Scholar
  9. 9.
    M. Criado, H. Eibl, and F.J. Barrantes, Effects of lipids on acetylcholine receptor. Essential need of cholesterol for maintenance of agonist-induced state transition in lipid vesicles, Biochemistry 21: 3622 (1982).PubMedCrossRefGoogle Scholar
  10. 10.
    L.L.M. van Deenen, Topology and dynamics of phospholipids in membranes, FEBS Lett. 123: 3 (1981).PubMedCrossRefGoogle Scholar
  11. 11.
    F.V. DeFeudis, A.N.K. Yusufi, L. Ossola, M. Maitre, P. Wolfe, G. Rebel, and P. Mandel, Antiserum to gangliosides inhibits (3H) GABA binding to a synaptosome-enriched fraction of rat cerebral cortex, Gen. Pharmacol. 11: 251 (1980).Google Scholar
  12. 12.
    G.E. De Medio, G. Trovarelli, A. Hamberger, and G. Porcellati, Synaptosomal phospholipid pool in rabbit brain and its effect on GABA uptake, Neurochem. Res., 5: 171 (1980).PubMedCrossRefGoogle Scholar
  13. 13.
    S. Fiszer de Plazas and E. de Robertis, Isolation of hydrophobic proteins binding amino acids. GABA binding in the rat cerebral cortex, J. Neurochem. 25: 547 (1975).CrossRefGoogle Scholar
  14. 14.
    M. Giesing, “Explantatkulturen des Nervensystems: Ein neues Modell für die Neurochemie. Bericht von der Regulation einiger Lipidbausteine”, Habilitationsschrift, Universität Bonn (1978).Google Scholar
  15. 15.
    M. Giesing and U. Gerken, The role of asymmetrically distributed phospholipids in the binding of gamma aminobutyric acid, in: “Basic and Clinical Aspects of Molecular Neurobiology”, A.M. Giuffrida Stella, G. Gombos, G. Benzi, and H.S. Bachelard, eds., Fondazione Internazionale Menarini, Milano, pp. 135 (1982).Google Scholar
  16. 16.
    M. Giesing and U. Gerken, The effects of carbamylcholine on extrasynaptic phosphatidylcholine biosynthesis in grey matter of cerebral cortex, in: “Phospholipid Metabolism in the Nervous System”, L.A. Horrocks, G.B. Ansell, and G. Porcellati, eds., Raven Press, New York, in press (1982).Google Scholar
  17. 17.
    M. Giesing, G. Neumann, H. Egge, and F. Zilliken, Lipid metabolism of developing central nervous tissues in organotypic cultures. I. Lipid distribution and fatty acid profiles of the medium for rat brain cortex in vitro, Nutr. Metabol. 19: 242 (1975).CrossRefGoogle Scholar
  18. 18.
    M. Giesing, B. Schmitz, B. Kempfle, H. Egge, and F. Zilliken, Effect of phosphoglycerolipids related to nutrition on GABA transport in cultured neurons, in: “New trends in nutrition, lipid research and cardiovascular diseases”, N.G.Bazan, J.M. Iacono, and R. Paoletti, eds., Alan R. Liss Inc., New York, pp. 45 (1981).Google Scholar
  19. 19.
    M. Giesing and F. Zilliken, Lipid metabolism of developing central nervous tissues in organotypic cultures. III. Ganglionic control of glycerolipids and fatty acids in cortex grey matter, Neurochem. Res. 5: 257 (1980).PubMedCrossRefGoogle Scholar
  20. 20.
    A Guidotti, G. Toffano, and E. Costa, An endogenous protein modulates the affinity of GABA and benzodiazepine receptors in rat brain, Nature 275: 553 (1978).PubMedCrossRefGoogle Scholar
  21. 21.
    J.N. Hawthorne and M.R. Pickard, Phospholipids in synaptic function, J. Neurochem. 32: 5 (1979).PubMedCrossRefGoogle Scholar
  22. 22.
    D.S. Heron, M. Israeli, M. Hershkovitz, D. Samuel, and M. Shinitzki, Lipid-induced modulation of opiate receptors in mouse brain membranes. Eur. J. Pharmacol. 72: 361 (1981).PubMedCrossRefGoogle Scholar
  23. 23.
    D.S. Heron, M. Shinitzki, M. Hershkovitz, and D. Samuel, Lipid fluidity markedly modulates the binding of serotonin to mouse brain membranes, Proc. Natl. Acad. Sci. U.S.A. 77: 7463 (1980).PubMedCrossRefGoogle Scholar
  24. 24.
    F. Hirata, J.F. Tallmann, R.C. Henneberry, P. Mallorga, W.J. Strittmatter, and J. Axelrod, Phospholipid methylation: A possible mechanism of signal transduction across biomembranes, in: “Membrane transport and neuroreceptors”, Alan R. Liss Inc., New York, pp. 383 (1981).Google Scholar
  25. 25.
    A.F. Horwitz, M.E. Hatten, and M.M. Burger, Membrane fatty acid replacement and their effect on growth and lectin-induced agglutinability, Proc. Natl. Acad. Sci. U.S.A., 71: 3115 (1974).PubMedCrossRefGoogle Scholar
  26. 26.
    R.J. Levkovitz, D. Mullikin, C. Wood, T. Goore, and C. Mukherjee, Regulation of Prostaglandin Receptors by Prostaglandins and Guanine Nucleotides in Frog Erythrocytes, J. Biol. Chem. 252: 5295 (1977).Google Scholar
  27. 27.
    F. Lembeck, A. Saria, and N. Mayer, Substance P: Model studies of its binding to phospholipids, Naunyn-Schmiedeberg’s Arch. Pharmacol. 306: 189 (1979).Google Scholar
  28. 28.
    K.G. Lloyd and K. Beaumont, Possible role of phospholipids in GABA receptor function in human and rat brain, Brain Res. Bull., 5: 285 (1980).CrossRefGoogle Scholar
  29. 29.
    M.D. Majewska, R. Manning, and G.Y. Sun, Effects of postdecapitative ischemia on arachidonate release from brain synaptosomes, Neurochem. Res. 6: 567 (1981).PubMedCrossRefGoogle Scholar
  30. 30.
    R.H. Ng and B.D. Howard, Inhibition of neurotoxic phospholipase A2 on synaptosomal uptake of γ-aminobutyric acid, J. Neurochem. 36: 310 (1981).PubMedCrossRefGoogle Scholar
  31. 31.
    A. Nistri and A. Constanti, Pharmacological characterization of different types of GABA and glutamate receptors in vertebrates and invertebrates, Progr. Neurobiol. 13: 117 (1979).CrossRefGoogle Scholar
  32. 32.
    R.W. Olsen, J.D. Bayless, and M. Ban, Potency of inhibitors for γ-aminobutyric acid uptake by mouse brain subcellular particles at 0°,Mol. Pharmacol. 11: 558 (1975).PubMedGoogle Scholar
  33. 33.
    J. Robert, P. Mandel, and G. Rebel, Neutral lipids and phospholipids from cultured astroblasts, J. Neurochem. 26: 771 (1976).PubMedCrossRefGoogle Scholar
  34. 34.
    E.J. Simon, Studies on membrane-bound opiate receptors, in: “Membrane mechanism of drugs and abuse”, Alan R. Liss Inc., New York, pp. 51 (1979).Google Scholar
  35. 35.
    H. Tamir, W. Brunner, D. Casper and M.R. Rapport, Enhancement by gangliosides of the binding of serotonin to serotonin binding proteins, J. Neurochem. 34: 1719 (1980).PubMedCrossRefGoogle Scholar
  36. 36.
    L. Thilo, H. Träuble, and P. Overath, Mechanistic interpretation of the influence of lipid phase transitions on transport functions, Biochemistry 16: 1283 (1977).PubMedCrossRefGoogle Scholar
  37. 37.
    G. Toffano, C. Aldinio, M. Balzano, A. Leon, and G. Savoini, Regulation of GABA receptor binding to synaptic plasma membrane of rat cerebral cortex: the role of endogenous phospholipids, Brain Res. 222: 95 (1981).PubMedCrossRefGoogle Scholar
  38. 38.
    L.T. Williams and R.J. Lefkovitz, Slowly reversible binding of catecholamine to a nucleotide-sensitive state of the ß-adrenergic receptor, J. Biol. Chem. 252: 7207 (1977).PubMedGoogle Scholar
  39. 39.
    J.R. Yandrasitz, R.M. Cohn, B. Masley, and D. Rowe, Evaluation of the binding of serotonin by isolated CNS acidic lipids, Neurochem.Res. 5: 465 (1980).PubMedCrossRefGoogle Scholar
  40. 40.
    Y. Yoneda and K. Kuriayama, Presence of a low molecular weight endogenous inhibitor on 3H-muscimol binding in synaptic membranes, Nature 285: 670 (1980).PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • Michael Giesing
    • 1
  1. 1.Nattermann Research LaboratoriesKöln 30GFR

Personalised recommendations