Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1988 Feb 1;106(2):487–503. doi: 10.1083/jcb.106.2.487

Neuron-glia cell adhesion molecule interacts with neurons and astroglia via different binding mechanisms

PMCID: PMC2114985  PMID: 2448316

Abstract

The neuron-glia cell adhesion molecule (Ng-CAM) is present in the central nervous system on postmitotic neurons and in the periphery on neurons and Schwann cells. It has been implicated in binding between neurons and between neurons and glia. To understand the molecular mechanisms of Ng-CAM binding, we analyzed the aggregation of chick Ng- CAM either immobilized on 0.5-micron beads (Covaspheres) or reconstituted into liposomes. The results were correlated with the binding of these particles to different types of cells as well as with cell-cell binding itself. Both Ng-CAM-Covaspheres and Ng-CAM liposomes individually self-aggregated, and antibodies against Ng-CAM strongly inhibited their aggregation; the rate of aggregation increased approximately with the square of the concentration of the beads or the liposomes. Much higher rates of aggregation were observed when the ratio of Ng-CAM to lipid in the liposome was increased. Radioiodinated Ng-CAM on Covaspheres and in liposomes bound both to neurons and to glial cells and in each case antibodies against Ng-CAM inhibited 50-90% of the binding. Control preparations of fibroblasts and meningeal cells did not exhibit significant binding. Adhesion between neurons and glia within and across species (chick and mouse) was explored in cellular assays after defining markers for each cell type, and optimal conditions of shear, temperature, and cell density. As previously noted using chick cells (Grumet, M., S. Hoffman, C.-M. Chuong, and G. M. Edelman. 1984 Proc. Natl. Acad. Sci. USA. 81:7989-7993), anti-Ng-CAM antibodies inhibited neuron-neuron and neuron-glia binding. In cross- species adhesion assays, binding of chick neurons to mouse astroglia and binding of mouse neurons to chick astroglia were both inhibited by anti-Ng-CAM antibodies. To identify whether the cellular ligands for Ng- CAM differed for neuron-neuron and neuron-glia binding, cells were preincubated with specific antibodies, the antibodies were removed by washing, and Ng-CAM-Covasphere binding was measured. Preincubation of neurons with anti-Ng-CAM antibodies inhibited Ng-CAM-Covasphere binding but similar preincubation of astroglial cells did not inhibit binding. In contrast, preincubation of astroglia with anti-astroglial cell antibodies inhibited binding to these cells but preincubation of neurons with these antibodies had no effect. Together with the data on Covaspheres and liposome aggregation, these findings suggested that Ng- CAM-Covaspheres bound to Ng-CAM on neurons but bound to different molecules on astroglia.(ABSTRACT TRUNCATED AT 400 WORDS)

Full Text

The Full Text of this article is available as a PDF (5.5 MB).

Selected References

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

  1. Adler R., Magistretti P. J., Hyndman A. G., Shoemaker W. J. Purification and cytochemical identification of neuronal and non-neuronal cells in chick embryo retina cultures. Dev Neurosci. 1982;5(1):27–39. doi: 10.1159/000112659. [DOI] [PubMed] [Google Scholar]
  2. Adler R., Manthorpe M., Varon S. Separation of neuronal and nonneuronal cells in monolayer cultures from chick embryo optic lobe. Dev Biol. 1979 Apr;69(2):424–435. doi: 10.1016/0012-1606(79)90302-6. [DOI] [PubMed] [Google Scholar]
  3. Antonicek H., Persohn E., Schachner M. Biochemical and functional characterization of a novel neuron-glia adhesion molecule that is involved in neuronal migration. J Cell Biol. 1987 Jun;104(6):1587–1595. doi: 10.1083/jcb.104.6.1587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bignami A., Eng L. F., Dahl D., Uyeda C. T. Localization of the glial fibrillary acidic protein in astrocytes by immunofluorescence. Brain Res. 1972 Aug 25;43(2):429–435. doi: 10.1016/0006-8993(72)90398-8. [DOI] [PubMed] [Google Scholar]
  5. Bock E., Richter-Landsberg C., Faissner A., Schachner M. Demonstration of immunochemical identity between the nerve growth factor-inducible large external (NILE) glycoprotein and the cell adhesion molecule L1. EMBO J. 1985 Nov;4(11):2765–2768. doi: 10.1002/j.1460-2075.1985.tb04001.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brackenbury R., Thiery J. P., Rutishauser U., Edelman G. M. Adhesion among neural cells of the chick embryo. I. An immunological assay for molecules involved in cell-cell binding. J Biol Chem. 1977 Oct 10;252(19):6835–6840. [PubMed] [Google Scholar]
  7. Chuong C. M., Crossin K. L., Edelman G. M. Sequential expression and differential function of multiple adhesion molecules during the formation of cerebellar cortical layers. J Cell Biol. 1987 Feb;104(2):331–342. doi: 10.1083/jcb.104.2.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chuong C. M., Edelman G. M. Alterations in neural cell adhesion molecules during development of different regions of the nervous system. J Neurosci. 1984 Sep;4(9):2354–2368. doi: 10.1523/JNEUROSCI.04-09-02354.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dahl D., Bignami A. Intermediate filaments in nervous tissue. Cell Muscle Motil. 1985;6:75–96. doi: 10.1007/978-1-4757-4723-2_4. [DOI] [PubMed] [Google Scholar]
  10. Daniloff J. K., Chuong C. M., Levi G., Edelman G. M. Differential distribution of cell adhesion molecules during histogenesis of the chick nervous system. J Neurosci. 1986 Mar;6(3):739–758. doi: 10.1523/JNEUROSCI.06-03-00739.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Daniloff J. K., Levi G., Grumet M., Rieger F., Edelman G. M. Altered expression of neuronal cell adhesion molecules induced by nerve injury and repair. J Cell Biol. 1986 Sep;103(3):929–945. doi: 10.1083/jcb.103.3.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dräger U. C., Edwards D. L., Barnstable C. J. Antibodies against filamentous components in discrete cell types of the mouse retina. J Neurosci. 1984 Aug;4(8):2025–2042. doi: 10.1523/JNEUROSCI.04-08-02025.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Edelman G. M. Cell adhesion molecules in the regulation of animal form and tissue pattern. Annu Rev Cell Biol. 1986;2:81–116. doi: 10.1146/annurev.cb.02.110186.000501. [DOI] [PubMed] [Google Scholar]
  14. Edelman G. M. Cell adhesion molecules. Science. 1983 Feb 4;219(4584):450–457. doi: 10.1126/science.6823544. [DOI] [PubMed] [Google Scholar]
  15. Edelman G. M. Expression of cell adhesion molecules during embryogenesis and regeneration. Exp Cell Res. 1985 Nov;161(1):1–16. doi: 10.1016/0014-4827(85)90485-9. [DOI] [PubMed] [Google Scholar]
  16. Fallon J. R. Neurite guidance by non-neuronal cells in culture: preferential outgrowth of peripheral neurites on glial as compared to nonglial cell surfaces. J Neurosci. 1985 Dec;5(12):3169–3177. doi: 10.1523/JNEUROSCI.05-12-03169.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fischer G., Künemund V., Schachner M. Neurite outgrowth patterns in cerebellar microexplant cultures are affected by antibodies to the cell surface glycoprotein L1. J Neurosci. 1986 Feb;6(2):605–612. doi: 10.1523/JNEUROSCI.06-02-00605.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Friedlander D. R., Grumet M., Edelman G. M. Nerve growth factor enhances expression of neuron-glia cell adhesion molecule in PC12 cells. J Cell Biol. 1986 Feb;102(2):413–419. doi: 10.1083/jcb.102.2.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Grumet M., Edelman G. M. Heterotypic binding between neuronal membrane vesicles and glial cells is mediated by a specific cell adhesion molecule. J Cell Biol. 1984 May;98(5):1746–1756. doi: 10.1083/jcb.98.5.1746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Grumet M., Hoffman S., Chuong C. M., Edelman G. M. Polypeptide components and binding functions of neuron-glia cell adhesion molecules. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7989–7993. doi: 10.1073/pnas.81.24.7989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Grumet M., Hoffman S., Crossin K. L., Edelman G. M. Cytotactin, an extracellular matrix protein of neural and non-neural tissues that mediates glia-neuron interaction. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8075–8079. doi: 10.1073/pnas.82.23.8075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Grumet M., Hoffman S., Edelman G. M. Two antigenically related neuronal cell adhesion molecules of different specificities mediate neuron-neuron and neuron-glia adhesion. Proc Natl Acad Sci U S A. 1984 Jan;81(1):267–271. doi: 10.1073/pnas.81.1.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Grumet M., Rutishauser U., Edelman G. M. Neural cell adhesion molecule is on embryonic muscle cells and mediates adhesion to nerve cells in vitro. Nature. 1982 Feb 25;295(5851):693–695. doi: 10.1038/295693a0. [DOI] [PubMed] [Google Scholar]
  24. Grumet M., Rutishauser U., Edelman G. M. Neuron-glia adhesion is inhibited by antibodies to neural determinants. Science. 1983 Oct 7;222(4619):60–62. doi: 10.1126/science.6194561. [DOI] [PubMed] [Google Scholar]
  25. Hanson G. R., Iversen P. L., Partlow L. M. Preparation and partial characterization of highly purified primary cultures of neurons and non-neuronal (glial) cells from embryonic chick cerebral hemispheres and several other regions of the nervous system. Brain Res. 1982 Apr;255(4):529–545. doi: 10.1016/0165-3806(82)90052-9. [DOI] [PubMed] [Google Scholar]
  26. Hatta K., Okada T. S., Takeichi M. A monoclonal antibody disrupting calcium-dependent cell-cell adhesion of brain tissues: possible role of its target antigen in animal pattern formation. Proc Natl Acad Sci U S A. 1985 May;82(9):2789–2793. doi: 10.1073/pnas.82.9.2789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Hatta K., Takagi S., Fujisawa H., Takeichi M. Spatial and temporal expression pattern of N-cadherin cell adhesion molecules correlated with morphogenetic processes of chicken embryos. Dev Biol. 1987 Mar;120(1):215–227. doi: 10.1016/0012-1606(87)90119-9. [DOI] [PubMed] [Google Scholar]
  28. Hoffman S., Chuong C. M., Edelman G. M. Evolutionary conservation of key structures and binding functions of neural cell adhesion molecules. Proc Natl Acad Sci U S A. 1984 Nov;81(21):6881–6885. doi: 10.1073/pnas.81.21.6881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hoffman S., Edelman G. M. A proteoglycan with HNK-1 antigenic determinants is a neuron-associated ligand for cytotactin. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2523–2527. doi: 10.1073/pnas.84.8.2523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Hoffman S., Edelman G. M. Kinetics of homophilic binding by embryonic and adult forms of the neural cell adhesion molecule. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5762–5766. doi: 10.1073/pnas.80.18.5762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Hoffman S., Friedlander D. R., Chuong C. M., Grumet M., Edelman G. M. Differential contributions of Ng-CAM and N-CAM to cell adhesion in different neural regions. J Cell Biol. 1986 Jul;103(1):145–158. doi: 10.1083/jcb.103.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Hoffman S., Sorkin B. C., White P. C., Brackenbury R., Mailhammer R., Rutishauser U., Cunningham B. A., Edelman G. M. Chemical characterization of a neural cell adhesion molecule purified from embryonic brain membranes. J Biol Chem. 1982 Jul 10;257(13):7720–7729. [PubMed] [Google Scholar]
  33. Keilhauer G., Faissner A., Schachner M. Differential inhibition of neurone-neurone, neurone-astrocyte and astrocyte-astrocyte adhesion by L1, L2 and N-CAM antibodies. Nature. 1985 Aug 22;316(6030):728–730. doi: 10.1038/316728a0. [DOI] [PubMed] [Google Scholar]
  34. Kücherer A., Faissner A., Schachner M. The novel carbohydrate epitope L3 is shared by some neural cell adhesion molecules. J Cell Biol. 1987 Jun;104(6):1597–1602. doi: 10.1083/jcb.104.6.1597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  37. Lemmon V., McLoon S. C. The appearance of an L1-like molecule in the chick primary visual pathway. J Neurosci. 1986 Oct;6(10):2987–2994. doi: 10.1523/JNEUROSCI.06-10-02987.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Lindner J., Guenther J., Nick H., Zinser G., Antonicek H., Schachner M., Monard D. Modulation of granule cell migration by a glia-derived protein. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4568–4571. doi: 10.1073/pnas.83.12.4568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lindner J., Zinser G., Werz W., Goridis C., Bizzini B., Schachner M. Experimental modification of postnatal cerebellar granule cell migration in vitro. Brain Res. 1986 Jul 9;377(2):298–304. doi: 10.1016/0006-8993(86)90872-3. [DOI] [PubMed] [Google Scholar]
  40. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  41. Mugnaini E., Forstronen P. F. Ultrastructural studies on the cerebellar histogenesis. I. Differentiation of granule cells and development of glomeruli in the chick embryo. Z Zellforsch Mikrosk Anat. 1967;77(1):115–143. doi: 10.1007/BF00336702. [DOI] [PubMed] [Google Scholar]
  42. Pigott R., Kelly J. S. Immunocytochemical and biochemical studies with the monoclonal antibody 69A1: similarities of the antigen with cell adhesion molecules L1, NILE and Ng-CAM. Brain Res. 1986 Sep;394(1):111–122. doi: 10.1016/0165-3806(86)90087-8. [DOI] [PubMed] [Google Scholar]
  43. Raff M. C., Mirsky R., Fields K. L., Lisak R. P., Dorfman S. H., Silberberg D. H., Gregson N. A., Leibowitz S., Kennedy M. C. Galactocerebroside is a specific cell-surface antigenic marker for oligodendrocytes in culture. Nature. 1978 Aug 24;274(5673):813–816. [PubMed] [Google Scholar]
  44. Rakic P. Neuron-glia relationship during granule cell migration in developing cerebellar cortex. A Golgi and electronmicroscopic study in Macacus Rhesus. J Comp Neurol. 1971 Mar;141(3):283–312. doi: 10.1002/cne.901410303. [DOI] [PubMed] [Google Scholar]
  45. Rathjen F. G., Rutishauser U. Comparison of two cell surface molecules involved in neural cell adhesion. EMBO J. 1984 Feb;3(2):461–465. doi: 10.1002/j.1460-2075.1984.tb01828.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rathjen F. G., Schachner M. Immunocytological and biochemical characterization of a new neuronal cell surface component (L1 antigen) which is involved in cell adhesion. EMBO J. 1984 Jan;3(1):1–10. doi: 10.1002/j.1460-2075.1984.tb01753.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Rathjen F. G., Wolff J. M., Frank R., Bonhoeffer F., Rutishauser U. Membrane glycoproteins involved in neurite fasciculation. J Cell Biol. 1987 Feb;104(2):343–353. doi: 10.1083/jcb.104.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Rieger F., Daniloff J. K., Pincon-Raymond M., Crossin K. L., Grumet M., Edelman G. M. Neuronal cell adhesion molecules and cytotactin are colocalized at the node of Ranvier. J Cell Biol. 1986 Aug;103(2):379–391. doi: 10.1083/jcb.103.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Sajovic P., Kouvelas E., Trenkner E. Probable identity of NILE glycoprotein and the high-molecular-weight component of L1 antigen. J Neurochem. 1986 Aug;47(2):541–546. doi: 10.1111/j.1471-4159.1986.tb04533.x. [DOI] [PubMed] [Google Scholar]
  50. Salton S. R., Shelanski M. L., Greene L. A. Biochemical properties of the nerve growth factor-inducible large external (NILE) glycoprotein. J Neurosci. 1983 Dec;3(12):2420–2430. doi: 10.1523/JNEUROSCI.03-12-02420.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Sensenbrenner M., Mandel P. Behaviour of neuroblasts in the presence of glial cells, fibroblasts and meningeal cells in culture. Exp Cell Res. 1974 Jul;87(1):159–167. doi: 10.1016/0014-4827(74)90538-2. [DOI] [PubMed] [Google Scholar]
  52. Silver J., Rutishauser U. Guidance of optic axons in vivo by a preformed adhesive pathway on neuroepithelial endfeet. Dev Biol. 1984 Dec;106(2):485–499. doi: 10.1016/0012-1606(84)90248-3. [DOI] [PubMed] [Google Scholar]
  53. Stallcup W. B., Beasley L. Involvement of the nerve growth factor-inducible large external glycoprotein (NILE) in neurite fasciculation in primary cultures of rat brain. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1276–1280. doi: 10.1073/pnas.82.4.1276. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Thiery J. P., Delouvée A., Grumet M., Edelman G. M. Initial appearance and regional distribution of the neuron-glia cell adhesion molecule in the chick embryo. J Cell Biol. 1985 Feb;100(2):442–456. doi: 10.1083/jcb.100.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Tholey G., Ledig M., Bloch S., Mandel P. Trypsinization of chick glial cells before seeding: effects on energy metabolism enzymes and glutamine synthetase. Neurochem Res. 1983 Oct;8(10):1233–1243. doi: 10.1007/BF00963994. [DOI] [PubMed] [Google Scholar]
  56. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Volk T., Geiger B. A-CAM: a 135-kD receptor of intercellular adherens junctions. I. Immunoelectron microscopic localization and biochemical studies. J Cell Biol. 1986 Oct;103(4):1441–1450. doi: 10.1083/jcb.103.4.1441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Volk T., Geiger B. A-CAM: a 135-kD receptor of intercellular adherens junctions. II. Antibody-mediated modulation of junction formation. J Cell Biol. 1986 Oct;103(4):1451–1464. doi: 10.1083/jcb.103.4.1451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Williams R. K., Goridis C., Akeson R. Individual neural cell types express immunologically distinct N-CAM forms. J Cell Biol. 1985 Jul;101(1):36–42. doi: 10.1083/jcb.101.1.36. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES