Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1988 Jan 1;106(1):213–223. doi: 10.1083/jcb.106.1.213

The L2/HNK-1 carbohydrate of neural cell adhesion molecules is involved in cell interactions

PMCID: PMC2114958  PMID: 2448311

Abstract

We investigated whether the L2/HNK-1 carbohydrate epitope, expressed by two unusual glycolipids and several neural adhesion molecules, including L1, neural cell adhesion molecule, J1, and the myelin- associated glycoprotein, is involved in adhesion. Monoclonal L2 antibodies, the L2/HNK-1-reactive, sulfate-3-glucuronyl residue carrying glycolipids (L2 glycolipid) and a tetrasaccharide derived from the L2 glycolipid (L2 tetrasaccharide) were added to microexplant cultures of early postnatal mouse cerebellum, and cell migration and process extension were monitored. On the substrate poly-D-lysine, Fab fragments of L2 antibodies, L2 glycolipid, and L2 tetrasaccharide inhibited outgrowth of astrocytic processes and migration of cell bodies, but only L2 glycolipid and L2 tetrasaccharide reduced neurite outgrowth. On laminin, L2 antibodies, L2 glycolipid, and L2 tetrasaccharide inhibited outgrowth of astrocytic processes. Additionally, L2 glycolipid and L2 tetrasaccharide inhibited cell migration and neurite outgrowth. Several negatively charged glycolipids, lipids, and saccharides were tested for control and found to have no effect on outgrowth patterns, except for sulfatide and heparin, which modified outgrowth patterns in a similar fashion as L2 glycolipid and L2 tetrasaccharide. On astrocytes none of the tested compounds interfered with explant outgrowth. In short-term adhesion assays L2 glycolipid, sulfatide, and heparin inhibited adhesion of neural cells to laminin. L2 glycolipid and sulfatide interfered with neuron to astrocyte and astrocyte to astrocyte adhesion, but not with neuron-neuron adhesion. The most straightforward interpretation of these observations is that the L2/HNK-1 carbohydrate and the sulfated carbohydrates, sulfatide and heparin, act as ligands in cell adhesion.

Full Text

The Full Text of this article is available as a PDF (4.7 MB).

Selected References

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

  1. Abo T., Balch C. M. A differentiation antigen of human NK and K cells identified by a monoclonal antibody (HNK-1). J Immunol. 1981 Sep;127(3):1024–1029. [PubMed] [Google Scholar]
  2. Ariga T., Kohriyama T., Freddo L., Latov N., Saito M., Kon K., Ando S., Suzuki M., Hemling M. E., Rinehart K. L., Jr Characterization of sulfated glucuronic acid containing glycolipids reacting with IgM M-proteins in patients with neuropathy. J Biol Chem. 1987 Jan 15;262(2):848–853. [PubMed] [Google Scholar]
  3. Baron-Van Evercooren A., Kleinman H. K., Ohno S., Marangos P., Schwartz J. P., Dubois-Dalcq M. E. Nerve growth factor, laminin, and fibronectin promote neurite growth in human fetal sensory ganglia cultures. J Neurosci Res. 1982;8(2-3):179–193. doi: 10.1002/jnr.490080208. [DOI] [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. Braun P. E., Frail D. E., Latov N. Myelin-associated glycoprotein is the antigen for a monoclonal IgM in polyneuropathy. J Neurochem. 1982 Nov;39(5):1261–1265. doi: 10.1111/j.1471-4159.1982.tb12563.x. [DOI] [PubMed] [Google Scholar]
  6. Calvo F. O., Ryan R. J. Inhibition of adenylyl cyclase activity in rat corpora luteal tissue by glycopeptides of human chorionic gonadotropin and the alpha-subunit of human chorionic gonadotropin. Biochemistry. 1985 Apr 9;24(8):1953–1959. doi: 10.1021/bi00329a023. [DOI] [PubMed] [Google Scholar]
  7. Chou D. K., Ilyas A. A., Evans J. E., Costello C., Quarles R. H., Jungalwala F. B. Structure of sulfated glucuronyl glycolipids in the nervous system reacting with HNK-1 antibody and some IgM paraproteins in neuropathy. J Biol Chem. 1986 Sep 5;261(25):11717–11725. [PubMed] [Google Scholar]
  8. Chou K. H., Ilyas A. A., Evans J. E., Quarles R. H., Jungalwala F. B. Structure of a glycolipid reacting with monoclonal IgM in neuropathy and with HNK-1. Biochem Biophys Res Commun. 1985 Apr 16;128(1):383–388. doi: 10.1016/0006-291x(85)91690-0. [DOI] [PubMed] [Google Scholar]
  9. Chou K. H., Nolan C. E., Jungalwala F. B. Composition and metabolism of gangliosides in rat peripheral nervous system during development. J Neurochem. 1982 Dec;39(6):1547–1558. doi: 10.1111/j.1471-4159.1982.tb07987.x. [DOI] [PubMed] [Google Scholar]
  10. Cole G. J., Glaser L. A heparin-binding domain from N-CAM is involved in neural cell-substratum adhesion. J Cell Biol. 1986 Feb;102(2):403–412. doi: 10.1083/jcb.102.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cole G. J., Loewy A., Cross N. V., Akeson R., Glaser L. Topographic localization of the heparin-binding domain of the neural cell adhesion molecule N-CAM. J Cell Biol. 1986 Nov;103(5):1739–1744. doi: 10.1083/jcb.103.5.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cole G. J., Loewy A., Glaser L. Neuronal cell-cell adhesion depends on interactions of N-CAM with heparin-like molecules. Nature. 1986 Apr 3;320(6061):445–447. doi: 10.1038/320445a0. [DOI] [PubMed] [Google Scholar]
  13. Cole G. J., Schachner M. Localization of the L2 monoclonal antibody binding site on chicken neural cell adhesion molecule (NCAM) and evidence for its role in NCAM-mediated cell adhesion. Neurosci Lett. 1987 Jul 22;78(2):227–232. doi: 10.1016/0304-3940(87)90638-0. [DOI] [PubMed] [Google Scholar]
  14. Edelman G. M. Cell adhesion and the molecular processes of morphogenesis. Annu Rev Biochem. 1985;54:135–169. doi: 10.1146/annurev.bi.54.070185.001031. [DOI] [PubMed] [Google Scholar]
  15. Edgar D., Timpl R., Thoenen H. The heparin-binding domain of laminin is responsible for its effects on neurite outgrowth and neuronal survival. EMBO J. 1984 Jul;3(7):1463–1468. doi: 10.1002/j.1460-2075.1984.tb01997.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Eng L. F., Vanderhaeghen J. J., Bignami A., Gerstl B. An acidic protein isolated from fibrous astrocytes. Brain Res. 1971 May 7;28(2):351–354. doi: 10.1016/0006-8993(71)90668-8. [DOI] [PubMed] [Google Scholar]
  17. Fischer G. Cultivation of mouse cerebellar cells in serum free, hormonally defined media: survival of neurons. Neurosci Lett. 1982 Mar 5;28(3):325–329. doi: 10.1016/0304-3940(82)90079-9. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Fischer G., Leutz A., Schachner M. Cultivation of immature astrocytes of mouse cerebellum in a serum-free, hormonally defined medium. Appearance of the mature astrocyte phenotype after addition of serum. Neurosci Lett. 1982 Apr 26;29(3):297–302. doi: 10.1016/0304-3940(82)90333-0. [DOI] [PubMed] [Google Scholar]
  20. Ilyas A. A., Dalakas M. C., Brady R. O., Quarles R. H. Sulfated glucuronyl glycolipids reacting with anti-myelin-associated glycoprotein monoclonal antibodies including IgM paraproteins in neuropathy: species distribution and partial characterization of epitopes. Brain Res. 1986 Oct 15;385(1):1–9. doi: 10.1016/0006-8993(86)91540-4. [DOI] [PubMed] [Google Scholar]
  21. Ilyas A. A., Quarles R. H., Brady R. O. The monoclonal antibody HNK-1 reacts with a human peripheral nerve ganglioside. Biochem Biophys Res Commun. 1984 Aug 16;122(3):1206–1211. doi: 10.1016/0006-291x(84)91220-8. [DOI] [PubMed] [Google Scholar]
  22. Ilyas A. A., Quarles R. H., MacIntosh T. D., Dobersen M. J., Trapp B. D., Dalakas M. C., Brady R. O. IgM in a human neuropathy related to paraproteinemia binds to a carbohydrate determinant in the myelin-associated glycoprotein and to a ganglioside. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1225–1229. doi: 10.1073/pnas.81.4.1225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Kruse J., Keilhauer G., Faissner A., Timpl R., Schachner M. The J1 glycoprotein--a novel nervous system cell adhesion molecule of the L2/HNK-1 family. Nature. 1985 Jul 11;316(6024):146–148. doi: 10.1038/316146a0. [DOI] [PubMed] [Google Scholar]
  25. Kruse J., Mailhammer R., Wernecke H., Faissner A., Sommer I., Goridis C., Schachner M. Neural cell adhesion molecules and myelin-associated glycoprotein share a common carbohydrate moiety recognized by monoclonal antibodies L2 and HNK-1. Nature. 1984 Sep 13;311(5982):153–155. doi: 10.1038/311153a0. [DOI] [PubMed] [Google Scholar]
  26. Liesi P., Dahl D., Vaheri A. Laminin is produced by early rat astrocytes in primary culture. J Cell Biol. 1983 Mar;96(3):920–924. doi: 10.1083/jcb.96.3.920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Manthorpe M., Engvall E., Ruoslahti E., Longo F. M., Davis G. E., Varon S. Laminin promotes neuritic regeneration from cultured peripheral and central neurons. J Cell Biol. 1983 Dec;97(6):1882–1890. doi: 10.1083/jcb.97.6.1882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McGarry R. C., Riopelle R. J., Roder J. C. Accelerated regenerative neurite formation by a neuronal surface epitope reactive with the monoclonal antibody, Leu 7. Neurosci Lett. 1985 May 14;56(2):95–100. doi: 10.1016/0304-3940(85)90113-2. [DOI] [PubMed] [Google Scholar]
  29. Noronha A. B., Ilyas A., Antonicek H., Schachner M., Quarles R. H. Molecular specificity of L2 monoclonal antibodies that bind to carbohydrate determinants of neural cell adhesion molecules and their resemblance to other monoclonal antibodies recognizing the myelin-associated glycoprotein. Brain Res. 1986 Oct 22;385(2):237–244. doi: 10.1016/0006-8993(86)91069-3. [DOI] [PubMed] [Google Scholar]
  30. PORTER R. R. The hydrolysis of rabbit y-globulin and antibodies with crystalline papain. Biochem J. 1959 Sep;73:119–126. doi: 10.1042/bj0730119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Poltorak M., Sadoul R., Keilhauer G., Landa C., Fahrig T., Schachner M. Myelin-associated glycoprotein, a member of the L2/HNK-1 family of neural cell adhesion molecules, is involved in neuron-oligodendrocyte and oligodendrocyte-oligodendrocyte interaction. J Cell Biol. 1987 Oct;105(4):1893–1899. doi: 10.1083/jcb.105.4.1893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Poltorak M., Steck A. J., Schachner M. Reactivity with neural cell adhesion molecules in sera from patients with demyelinating diseases. Neurosci Lett. 1986 Apr 11;65(2):199–203. doi: 10.1016/0304-3940(86)90304-6. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Riopelle R. J., McGarry R. C., Roder J. C. Adhesion properties of a neuronal epitope recognized by the monoclonal antibody HNK-1. Brain Res. 1986 Mar 5;367(1-2):20–25. doi: 10.1016/0006-8993(86)91573-8. [DOI] [PubMed] [Google Scholar]
  35. Roberts D. D., Rao C. N., Liotta L. A., Gralnick H. R., Ginsburg V. Comparison of the specificities of laminin, thrombospondin, and von Willebrand factor for binding to sulfated glycolipids. J Biol Chem. 1986 May 25;261(15):6872–6877. [PubMed] [Google Scholar]
  36. Roberts D. D., Rao C. N., Magnani J. L., Spitalnik S. L., Liotta L. A., Ginsburg V. Laminin binds specifically to sulfated glycolipids. Proc Natl Acad Sci U S A. 1985 Mar;82(5):1306–1310. doi: 10.1073/pnas.82.5.1306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rogers S. L., Letourneau P. C., Palm S. L., McCarthy J., Furcht L. T. Neurite extension by peripheral and central nervous system neurons in response to substratum-bound fibronectin and laminin. Dev Biol. 1983 Jul;98(1):212–220. doi: 10.1016/0012-1606(83)90350-0. [DOI] [PubMed] [Google Scholar]
  38. Sanes J. R., Schachner M., Covault J. Expression of several adhesive macromolecules (N-CAM, L1, J1, NILE, uvomorulin, laminin, fibronectin, and a heparan sulfate proteoglycan) in embryonic, adult, and denervated adult skeletal muscle. J Cell Biol. 1986 Feb;102(2):420–431. doi: 10.1083/jcb.102.2.420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Schachner M., Faissner A., Kruse J., Lindner J., Meier D. H., Rathjen F. G., Wernecke H. Cell-type specificity and developmental expression of neural cell-surface components involved in cell interactions and of structurally related molecules. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 2):557–568. doi: 10.1101/sqb.1983.048.01.060. [DOI] [PubMed] [Google Scholar]
  40. Schnitzer J., Franke W. W., Schachner M. Immunocytochemical demonstration of vimentin in astrocytes and ependymal cells of developing and adult mouse nervous system. J Cell Biol. 1981 Aug;90(2):435–447. doi: 10.1083/jcb.90.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schnitzer J., Schachner M. Expression of Thy-1, H-2, and NS-4 cell surface antigens and tetanus toxin receptors in early postnatal and adult mouse cerebellum. J Neuroimmunol. 1981 Dec;1(4):429–456. doi: 10.1016/0165-5728(81)90022-9. [DOI] [PubMed] [Google Scholar]
  42. Steck A. J., Murray N., Meier C., Page N., Perruisseau G. Demyelinating neuropathy and monoclonal IgM antibody to myelin-associated glycoprotein. Neurology. 1983 Jan;33(1):19–23. doi: 10.1212/wnl.33.1.19. [DOI] [PubMed] [Google Scholar]
  43. Yamada K. M., Kennedy D. W., Grotendorst G. R., Momoi T. Glycolipids: receptors for fibronectin? J Cell Physiol. 1981 Nov;109(2):343–351. doi: 10.1002/jcp.1041090218. [DOI] [PubMed] [Google Scholar]

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

RESOURCES