Search tips
Search criteria 


Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1994 May 1; 125(3): 669–680.
PMCID: PMC2119998

The neuronal chondroitin sulfate proteoglycan neurocan binds to the neural cell adhesion molecules Ng-CAM/L1/NILE and N-CAM, and inhibits neuronal adhesion and neurite outgrowth


We have previously shown that aggregation of microbeads coated with N- CAM and Ng-CAM is inhibited by incubation with soluble neurocan, a chondroitin sulfate proteoglycan of brain, suggesting that neurocan binds to these cell adhesion molecules (Grumet, M., A. Flaccus, and R. U. Margolis. 1993. J. Cell Biol. 120:815). To investigate these interactions more directly, we have tested binding of soluble 125I- neurocan to microwells coated with different glycoproteins. Neurocan bound at high levels to Ng-CAM and N-CAM, but little or no binding was detected to myelin-associated glycoprotein, EGF receptor, fibronectin, laminin, and collagen IV. The binding to Ng-CAM and N-CAM was saturable and in each case Scatchard plots indicated a high affinity binding site with a dissociation constant of approximately 1 nM. Binding was significantly reduced after treatment of neurocan with chondroitinase, and free chondroitin sulfate inhibited binding of neurocan to Ng-CAM and N-CAM. These results indicate a role for chondroitin sulfate in this process, although the core glycoprotein also has binding activity. The COOH-terminal half of neurocan was shown to have binding properties essentially identical to those of the full-length proteoglycan. To study the potential biological functions of neurocan, its effects on neuronal adhesion and neurite growth were analyzed. When neurons were incubated on dishes coated with different combinations of neurocan and Ng-CAM, neuronal adhesion and neurite extension were inhibited. Experiments using anti-Ng-CAM antibodies as a substrate also indicate that neurocan has a direct inhibitory effect on neuronal adhesion and neurite growth. Immunoperoxidase staining of tissue sections showed that neurocan, Ng-CAM, and N-CAM are all present at highest concentration in the molecular layer and fiber tracts of developing cerebellum. The overlapping localization in vivo, the molecular binding studies, and the striking effects on neuronal adhesion and neurite growth support the view that neurocan may modulate neuronal adhesion and neurite growth during development by binding to neural cell adhesion molecules.

Full Text

The Full Text of this article is available as a PDF (2.6M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Becker JW, Erickson HP, Hoffman S, Cunningham BA, Edelman GM. Topology of cell adhesion molecules. Proc Natl Acad Sci U S A. 1989 Feb;86(3):1088–1092. [PubMed]
  • Brackenbury R, Rutishauser U, Edelman GM. Distinct calcium-independent and calcium-dependent adhesion systems of chicken embryo cells. Proc Natl Acad Sci U S A. 1981 Jan;78(1):387–391. [PubMed]
  • Brittis PA, Canning DR, Silver J. Chondroitin sulfate as a regulator of neuronal patterning in the retina. Science. 1992 Feb 7;255(5045):733–736. [PubMed]
  • Burgoon MP, Grumet M, Mauro V, Edelman GM, Cunningham BA. Structure of the chicken neuron-glia cell adhesion molecule, Ng-CAM: origin of the polypeptides and relation to the Ig superfamily. J Cell Biol. 1991 Mar;112(5):1017–1029. [PMC free article] [PubMed]
  • Calof AL, Lander AD. Relationship between neuronal migration and cell-substratum adhesion: laminin and merosin promote olfactory neuronal migration but are anti-adhesive. J Cell Biol. 1991 Nov;115(3):779–794. [PMC free article] [PubMed]
  • Crossin KL, Prieto AL, Hoffman S, Jones FS, Friedlander DR. Expression of adhesion molecules and the establishment of boundaries during embryonic and neural development. Exp Neurol. 1990 Jul;109(1):6–18. [PubMed]
  • Doherty P, Walsh FS. Cell adhesion molecules, second messengers and axonal growth. Curr Opin Neurobiol. 1992 Oct;2(5):595–601. [PubMed]
  • Edelman GM, Crossin KL. Cell adhesion molecules: implications for a molecular histology. Annu Rev Biochem. 1991;60:155–190. [PubMed]
  • Faltz LL, Reddi AH, Hascall GK, Martin D, Pita JC, Hascall VC. Characteristics of proteoglycans extracted from the Swarm rat chondrosarcoma with associative solvents. J Biol Chem. 1979 Feb 25;254(4):1375–1380. [PubMed]
  • Friedlander DR, Hoffman S, Edelman GM. Functional mapping of cytotactin: proteolytic fragments active in cell-substrate adhesion. J Cell Biol. 1988 Dec;107(6 Pt 1):2329–2340. [PMC free article] [PubMed]
  • Grumet M. Structure, expression, and function of Ng-CAM, a member of the immunoglobulin superfamily involved in neuron-neuron and neuron-glia adhesion. J Neurosci Res. 1992 Jan;31(1):1–13. [PubMed]
  • Grumet M, Edelman GM. 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. [PMC free article] [PubMed]
  • Grumet M, Edelman GM. Neuron-glia cell adhesion molecule interacts with neurons and astroglia via different binding mechanisms. J Cell Biol. 1988 Feb;106(2):487–503. [PMC free article] [PubMed]
  • Grumet M, Flaccus A, Margolis RU. Functional characterization of chondroitin sulfate proteoglycans of brain: interactions with neurons and neural cell adhesion molecules. J Cell Biol. 1993 Feb;120(3):815–824. [PMC free article] [PubMed]
  • Grumet M, Hoffman S, Chuong CM, Edelman GM. Polypeptide components and binding functions of neuron-glia cell adhesion molecules. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7989–7993. [PubMed]
  • Grumet M, Hoffman S, Edelman GM. 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. [PubMed]
  • Hoffman S, Edelman GM. 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. [PubMed]
  • Hynes RO, Lander AD. Contact and adhesive specificities in the associations, migrations, and targeting of cells and axons. Cell. 1992 Jan 24;68(2):303–322. [PubMed]
  • Katoh-Semba R, Oohira A. Core proteins of soluble chondroitin sulfate proteoglycans purified from the rat brain block the cell cycle of PC12D cells. J Cell Physiol. 1993 Jul;156(1):17–23. [PubMed]
  • Kiang WL, Margolis RU, Margolis RK. Fractionation and properties of a chondroitin sulfate proteoglycan and the soluble glycoproteins of brain. J Biol Chem. 1981 Oct 25;256(20):10529–10537. [PubMed]
  • Krueger RC, Jr, Hennig AK, Schwartz NB. Two immunologically and developmentally distinct chondroitin sulfate proteolglycans in embryonic chick brain. J Biol Chem. 1992 Jun 15;267(17):12149–12161. [PubMed]
  • 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. [PubMed]
  • Lax I, Mitra AK, Ravera C, Hurwitz DR, Rubinstein M, Ullrich A, Stroud RM, Schlessinger J. Epidermal growth factor (EGF) induces oligomerization of soluble, extracellular, ligand-binding domain of EGF receptor. A low resolution projection structure of the ligand-binding domain. J Biol Chem. 1991 Jul 25;266(21):13828–13833. [PubMed]
  • Lemmon V, Burden SM, Payne HR, Elmslie GJ, Hlavin ML. Neurite growth on different substrates: permissive versus instructive influences and the role of adhesive strength. J Neurosci. 1992 Mar;12(3):818–826. [PubMed]
  • Lochter A, Vaughan L, Kaplony A, Prochiantz A, Schachner M, Faissner A. J1/tenascin in substrate-bound and soluble form displays contrary effects on neurite outgrowth. J Cell Biol. 1991 Jun;113(5):1159–1171. [PMC free article] [PubMed]
  • Margolis RK, Goossen B, Tekotte H, Hilgenberg L, Margolis RU. Effects of beta-xylosides on proteoglycan biosynthesis and morphology of PC12 pheochromocytoma cells and primary cultures of rat cerebellum. J Cell Sci. 1991 Jun;99(Pt 2):237–246. [PubMed]
  • Margolis RK, Margolis RU. Nervous tissue proteoglycans. Experientia. 1993 May 15;49(5):429–446. [PubMed]
  • Maurel P, Rauch U, Flad M, Margolis RK, Margolis RU. Phosphacan, a chondroitin sulfate proteoglycan of brain that interacts with neurons and neural cell-adhesion molecules, is an extracellular variant of a receptor-type protein tyrosine phosphatase. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2512–2516. [PubMed]
  • Moos M, Tacke R, Scherer H, Teplow D, Früh K, Schachner M. Neural adhesion molecule L1 as a member of the immunoglobulin superfamily with binding domains similar to fibronectin. Nature. 1988 Aug 25;334(6184):701–703. [PubMed]
  • Munson PJ, Rodbard D. Ligand: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem. 1980 Sep 1;107(1):220–239. [PubMed]
  • Oakley RA, Tosney KW. Peanut agglutinin and chondroitin-6-sulfate are molecular markers for tissues that act as barriers to axon advance in the avian embryo. Dev Biol. 1991 Sep;147(1):187–206. [PubMed]
  • Oh-eda M, Hasegawa M, Hattori K, Kuboniwa H, Kojima T, Orita T, Tomonou K, Yamazaki T, Ochi N. O-linked sugar chain of human granulocyte colony-stimulating factor protects it against polymerization and denaturation allowing it to retain its biological activity. J Biol Chem. 1990 Jul 15;265(20):11432–11435. [PubMed]
  • Pedraza L, Owens GC, Green LA, Salzer JL. The myelin-associated glycoproteins: membrane disposition, evidence of a novel disulfide linkage between immunoglobulin-like domains, and posttranslational palmitylation. J Cell Biol. 1990 Dec;111(6 Pt 1):2651–2661. [PMC free article] [PubMed]
  • Perris R, Krotoski D, Lallier T, Domingo C, Sorrell JM, Bronner-Fraser M. Spatial and temporal changes in the distribution of proteoglycans during avian neural crest development. Development. 1991 Feb;111(2):583–599. [PubMed]
  • Pindzola RR, Doller C, Silver J. Putative inhibitory extracellular matrix molecules at the dorsal root entry zone of the spinal cord during development and after root and sciatic nerve lesions. Dev Biol. 1993 Mar;156(1):34–48. [PubMed]
  • Pollerberg EG, Sadoul R, Goridis C, Schachner M. Selective expression of the 180-kD component of the neural cell adhesion molecule N-CAM during development. J Cell Biol. 1985 Nov;101(5 Pt 1):1921–1929. [PMC free article] [PubMed]
  • Prince JT, Alberti L, Healy PA, Nauman SJ, Stallcup WB. Molecular cloning of NILE glycoprotein and evidence for its continued expression in mature rat CNS. J Neurosci Res. 1991 Nov;30(3):567–581. [PubMed]
  • Rathjen FG, Rutishauser U. Comparison of two cell surface molecules involved in neural cell adhesion. EMBO J. 1984 Feb;3(2):461–465. [PubMed]
  • Rauch U, Gao P, Janetzko A, Flaccus A, Hilgenberg L, Tekotte H, Margolis RK, Margolis RU. Isolation and characterization of developmentally regulated chondroitin sulfate and chondroitin/keratan sulfate proteoglycans of brain identified with monoclonal antibodies. J Biol Chem. 1991 Aug 5;266(22):14785–14801. [PubMed]
  • Rauch U, Karthikeyan L, Maurel P, Margolis RU, Margolis RK. Cloning and primary structure of neurocan, a developmentally regulated, aggregating chondroitin sulfate proteoglycan of brain. J Biol Chem. 1992 Sep 25;267(27):19536–19547. [PubMed]
  • Reichardt LF, Tomaselli KJ. Extracellular matrix molecules and their receptors: functions in neural development. Annu Rev Neurosci. 1991;14:531–570. [PMC free article] [PubMed]
  • Ruoslahti E. Proteoglycans in cell regulation. J Biol Chem. 1989 Aug 15;264(23):13369–13372. [PubMed]
  • Schlessinger J. The epidermal growth factor receptor as a multifunctional allosteric protein. Biochemistry. 1988 May 3;27(9):3119–3123. [PubMed]
  • Schuch U, Lohse MJ, Schachner M. Neural cell adhesion molecules influence second messenger systems. Neuron. 1989 Jul;3(1):13–20. [PubMed]
  • Schwab ME, Kapfhammer JP, Bandtlow CE. Inhibitors of neurite growth. Annu Rev Neurosci. 1993;16:565–595. [PubMed]
  • Snow DM, Lemmon V, Carrino DA, Caplan AI, Silver J. Sulfated proteoglycans in astroglial barriers inhibit neurite outgrowth in vitro. Exp Neurol. 1990 Jul;109(1):111–130. [PubMed]
  • Snow DM, Steindler DA, Silver J. Molecular and cellular characterization of the glial roof plate of the spinal cord and optic tectum: a possible role for a proteoglycan in the development of an axon barrier. Dev Biol. 1990 Apr;138(2):359–376. [PubMed]
  • Snow DM, Letourneau PC. Neurite outgrowth on a step gradient of chondroitin sulfate proteoglycan (CS-PG). J Neurobiol. 1992 Apr;23(3):322–336. [PubMed]
  • Snow DM, Watanabe M, Letourneau PC, Silver J. A chondroitin sulfate proteoglycan may influence the direction of retinal ganglion cell outgrowth. Development. 1991 Dec;113(4):1473–1485. [PubMed]
  • Sonderegger P, Rathjen FG. Regulation of axonal growth in the vertebrate nervous system by interactions between glycoproteins belonging to two subgroups of the immunoglobulin superfamily. J Cell Biol. 1992 Dec;119(6):1387–1394. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Von Bohlen Und Halbach Friedrich, Taylor Joanne, Schachner Melitta. Cell Type-specific Effects of the Neural Adhesion Molecules L1 and N-CAM on Diverse Second Messenger Systems. Eur J Neurosci. 1992;4(10):896–909. [PubMed]
  • Wight TN, Kinsella MG, Qwarnström EE. The role of proteoglycans in cell adhesion, migration and proliferation. Curr Opin Cell Biol. 1992 Oct;4(5):793–801. [PubMed]
  • Zisch AH, D'Alessandri L, Ranscht B, Falchetto R, Winterhalter KH, Vaughan L. Neuronal cell adhesion molecule contactin/F11 binds to tenascin via its immunoglobulin-like domains. J Cell Biol. 1992 Oct;119(1):203–213. [PMC free article] [PubMed]

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