Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Apr 2;125(2):427–436. doi: 10.1083/jcb.125.2.427

Expression of NCAM containing VASE in neurons can account for a developmental loss in their neurite outgrowth response to NCAM in a cellular substratum

PMCID: PMC2120034  PMID: 8163558

Abstract

Binding of the neural cell adhesion molecule (NCAM) in neurons to NCAM on non-neuronal cells can stimulate axonal growth. A developmentally regulated loss of this response is associated with the insertion of 10 amino acids (called VASE) into the fourth Ig domain in up to 50% of the NCAM receptors in neurons. In the present study we have transfected PC12 cells with the major neuronal isoforms of human NCAM and tested cells expressing these isoforms for their ability to respond to NCAM in a cellular substratum. Whereas both the 140- and 180-kD isoforms of NCAM can act as functional receptors for neurite outgrowth, the presence of the VASE sequence in a minority of the receptors specifically inhibited this response. A synthetic peptide containing the VASE sequence inhibits neurite outgrowth from PC12 cells and primary neurons stimulated by NCAM. The same peptide has no effect on integrin dependent neurite outgrowth or neurite outgrowth stimulated by N-cadherin or L1. We discuss the possibility that the VASE peptide inhibits the NCAM response by preventing NCAM from binding to the FGF receptor in the plasma membrane.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

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

  1. Barton C. H., Dickson G., Gower H. J., Rowett L. H., Putt W., Elsom V., Moore S. E., Goridis C., Walsh F. S. Complete sequence and in vitro expression of a tissue-specific phosphatidylinositol-linked N-CAM isoform from skeletal muscle. Development. 1988 Sep;104(1):165–173. doi: 10.1242/dev.104.1.165. [DOI] [PubMed] [Google Scholar]
  2. Bixby J. L., Pratt R. S., Lilien J., Reichardt L. F. Neurite outgrowth on muscle cell surfaces involves extracellular matrix receptors as well as Ca2+-dependent and -independent cell adhesion molecules. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2555–2559. doi: 10.1073/pnas.84.8.2555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blaschuk O. W., Sullivan R., David S., Pouliot Y. Identification of a cadherin cell adhesion recognition sequence. Dev Biol. 1990 May;139(1):227–229. doi: 10.1016/0012-1606(90)90290-y. [DOI] [PubMed] [Google Scholar]
  4. Byers S., Amaya E., Munro S., Blaschuk O. Fibroblast growth factor receptors contain a conserved HAV region common to cadherins and influenza strain A hemagglutinins: a role in protein-protein interactions? Dev Biol. 1992 Aug;152(2):411–414. doi: 10.1016/0012-1606(92)90149-b. [DOI] [PubMed] [Google Scholar]
  5. Doherty P., Ashton S. V., Moore S. E., Walsh F. S. Morphoregulatory activities of NCAM and N-cadherin can be accounted for by G protein-dependent activation of L- and N-type neuronal Ca2+ channels. Cell. 1991 Oct 4;67(1):21–33. doi: 10.1016/0092-8674(91)90569-k. [DOI] [PubMed] [Google Scholar]
  6. Doherty P., Barton C. H., Dickson G., Seaton P., Rowett L. H., Moore S. E., Gower H. J., Walsh F. S. Neuronal process outgrowth of human sensory neurons on monolayers of cells transfected with cDNAs for five human N-CAM isoforms. J Cell Biol. 1989 Aug;109(2):789–798. doi: 10.1083/jcb.109.2.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Doherty P., Cohen J., Walsh F. S. Neurite outgrowth in response to transfected N-CAM changes during development and is modulated by polysialic acid. Neuron. 1990 Aug;5(2):209–219. doi: 10.1016/0896-6273(90)90310-c. [DOI] [PubMed] [Google Scholar]
  8. Doherty P., Fruns M., Seaton P., Dickson G., Barton C. H., Sears T. A., Walsh F. S. A threshold effect of the major isoforms of NCAM on neurite outgrowth. Nature. 1990 Feb 1;343(6257):464–466. doi: 10.1038/343464a0. [DOI] [PubMed] [Google Scholar]
  9. Doherty P., Mann D. A., Walsh F. S. Comparison of the effects of NGF, activators of protein kinase C, and a calcium ionophore on the expression of Thy-1 and N-CAM in PC12 cell cultures. J Cell Biol. 1988 Jul;107(1):333–340. doi: 10.1083/jcb.107.1.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Doherty P., Moolenaar C. E., Ashton S. V., Michalides R. J., Walsh F. S. The VASE exon downregulates the neurite growth-promoting activity of NCAM 140. Nature. 1992 Apr 30;356(6372):791–793. doi: 10.1038/356791a0. [DOI] [PubMed] [Google Scholar]
  11. Doherty P., Rimon G., Mann D. A., Walsh F. S. Alternative splicing of the cytoplasmic domain of neural cell adhesion molecule alters its ability to act as a substrate for neurite outgrowth. J Neurochem. 1992 Jun;58(6):2338–2341. doi: 10.1111/j.1471-4159.1992.tb10984.x. [DOI] [PubMed] [Google Scholar]
  12. Doherty P., Rowett L. H., Moore S. E., Mann D. A., Walsh F. S. Neurite outgrowth in response to transfected N-CAM and N-cadherin reveals fundamental differences in neuronal responsiveness to CAMs. Neuron. 1991 Feb;6(2):247–258. doi: 10.1016/0896-6273(91)90360-c. [DOI] [PubMed] [Google Scholar]
  13. Doherty P., Skaper S. D., Moore S. E., Leon A., Walsh F. S. A developmentally regulated switch in neuronal responsiveness to NCAM and N-cadherin in the rat hippocampus. Development. 1992 Jul;115(3):885–892. doi: 10.1242/dev.115.3.885. [DOI] [PubMed] [Google Scholar]
  14. Doherty P., Walsh F. S. Cell adhesion molecules, second messengers and axonal growth. Curr Opin Neurobiol. 1992 Oct;2(5):595–601. doi: 10.1016/0959-4388(92)90024-f. [DOI] [PubMed] [Google Scholar]
  15. Doherty P., Walsh F. S. Neurite guidance molecules. Curr Opin Cell Biol. 1989 Dec;1(6):1102–1106. doi: 10.1016/s0955-0674(89)80057-2. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Fraser S. E., Carhart M. S., Murray B. A., Chuong C. M., Edelman G. M. Alterations in the Xenopus retinotectal projection by antibodies to Xenopus N-CAM. Dev Biol. 1988 Sep;129(1):217–230. doi: 10.1016/0012-1606(88)90176-5. [DOI] [PubMed] [Google Scholar]
  18. Gower H. J., Barton C. H., Elsom V. L., Thompson J., Moore S. E., Dickson G., Walsh F. S. Alternative splicing generates a secreted form of N-CAM in muscle and brain. Cell. 1988 Dec 23;55(6):955–964. doi: 10.1016/0092-8674(88)90241-3. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Johnson D. E., Lee P. L., Lu J., Williams L. T. Diverse forms of a receptor for acidic and basic fibroblast growth factors. Mol Cell Biol. 1990 Sep;10(9):4728–4736. doi: 10.1128/mcb.10.9.4728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Landmesser L., Dahm L., Schultz K., Rutishauser U. Distinct roles for adhesion molecules during innervation of embryonic chick muscle. Dev Biol. 1988 Dec;130(2):645–670. doi: 10.1016/0012-1606(88)90358-2. [DOI] [PubMed] [Google Scholar]
  22. Landmesser L., Dahm L., Tang J. C., Rutishauser U. Polysialic acid as a regulator of intramuscular nerve branching during embryonic development. Neuron. 1990 May;4(5):655–667. doi: 10.1016/0896-6273(90)90193-j. [DOI] [PubMed] [Google Scholar]
  23. Liu L., Haines S., Shew R., Akeson R. A. Axon growth is enhanced by NCAM lacking the VASE exon when expressed in either the growth substrate or the growing axon. J Neurosci Res. 1993 Jun 15;35(3):327–345. doi: 10.1002/jnr.490350313. [DOI] [PubMed] [Google Scholar]
  24. Mann D. A., Doherty P., Walsh F. S. Increased intracellular cyclic AMP differentially modulates nerve growth factor induction of three neuronal recognition molecules involved in neurite outgrowth. J Neurochem. 1989 Nov;53(5):1581–1588. doi: 10.1111/j.1471-4159.1989.tb08555.x. [DOI] [PubMed] [Google Scholar]
  25. Matsunaga M., Hatta K., Nagafuchi A., Takeichi M. Guidance of optic nerve fibres by N-cadherin adhesion molecules. Nature. 1988 Jul 7;334(6177):62–64. doi: 10.1038/334062a0. [DOI] [PubMed] [Google Scholar]
  26. Mayford M., Barzilai A., Keller F., Schacher S., Kandel E. R. Modulation of an NCAM-related adhesion molecule with long-term synaptic plasticity in Aplysia. Science. 1992 May 1;256(5057):638–644. doi: 10.1126/science.1585176. [DOI] [PubMed] [Google Scholar]
  27. Moore S. E., Thompson J., Kirkness V., Dickson J. G., Walsh F. S. Skeletal muscle neural cell adhesion molecule (N-CAM): changes in protein and mRNA species during myogenesis of muscle cell lines. J Cell Biol. 1987 Sep;105(3):1377–1386. doi: 10.1083/jcb.105.3.1377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Neugebauer K. M., Tomaselli K. J., Lilien J., Reichardt L. F. N-cadherin, NCAM, and integrins promote retinal neurite outgrowth on astrocytes in vitro. J Cell Biol. 1988 Sep;107(3):1177–1187. doi: 10.1083/jcb.107.3.1177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Patel K., Moore S. E., Dickson G., Rossell R. J., Beverley P. C., Kemshead J. T., Walsh F. S. Neural cell adhesion molecule (NCAM) is the antigen recognized by monoclonal antibodies of similar specificity in small-cell lung carcinoma and neuroblastoma. Int J Cancer. 1989 Oct 15;44(4):573–578. doi: 10.1002/ijc.2910440402. [DOI] [PubMed] [Google Scholar]
  30. Rao Y., Wu X. F., Gariepy J., Rutishauser U., Siu C. H. Identification of a peptide sequence involved in homophilic binding in the neural cell adhesion molecule NCAM. J Cell Biol. 1992 Aug;118(4):937–949. doi: 10.1083/jcb.118.4.937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Reichardt L. F., Tomaselli K. J. Extracellular matrix molecules and their receptors: functions in neural development. Annu Rev Neurosci. 1991;14:531–570. doi: 10.1146/annurev.ne.14.030191.002531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Saffell J. L., Walsh F. S., Doherty P. Direct activation of second messenger pathways mimics cell adhesion molecule-dependent neurite outgrowth. J Cell Biol. 1992 Aug;118(3):663–670. doi: 10.1083/jcb.118.3.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schwab M. E. Myelin-associated inhibitors of neurite growth and regeneration in the CNS. Trends Neurosci. 1990 Nov;13(11):452–456. doi: 10.1016/0166-2236(90)90098-u. [DOI] [PubMed] [Google Scholar]
  34. Seilheimer B., Schachner M. Studies of adhesion molecules mediating interactions between cells of peripheral nervous system indicate a major role for L1 in mediating sensory neuron growth on Schwann cells in culture. J Cell Biol. 1988 Jul;107(1):341–351. doi: 10.1083/jcb.107.1.341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Small S. J., Akeson R. Expression of the unique NCAM VASE exon is independently regulated in distinct tissues during development. J Cell Biol. 1990 Nov;111(5 Pt 1):2089–2096. doi: 10.1083/jcb.111.5.2089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Small S. J., Haines S. L., Akeson R. A. Polypeptide variation in an N-CAM extracellular immunoglobulin-like fold is developmentally regulated through alternative splicing. Neuron. 1988 Dec;1(10):1007–1017. doi: 10.1016/0896-6273(88)90158-4. [DOI] [PubMed] [Google Scholar]
  37. Takeichi M. Cadherin cell adhesion receptors as a morphogenetic regulator. Science. 1991 Mar 22;251(5000):1451–1455. doi: 10.1126/science.2006419. [DOI] [PubMed] [Google Scholar]
  38. Tiveron M. C., Barboni E., Pliego Rivero F. B., Gormley A. M., Seeley P. J., Grosveld F., Morris R. Selective inhibition of neurite outgrowth on mature astrocytes by Thy-1 glycoprotein. Nature. 1992 Feb 20;355(6362):745–748. doi: 10.1038/355745a0. [DOI] [PubMed] [Google Scholar]
  39. Vidal-Sanz M., Bray G. M., Villegas-Pérez M. P., Thanos S., Aguayo A. J. Axonal regeneration and synapse formation in the superior colliculus by retinal ganglion cells in the adult rat. J Neurosci. 1987 Sep;7(9):2894–2909. doi: 10.1523/JNEUROSCI.07-09-02894.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Walicke P. A. Novel neurotrophic factors, receptors, and oncogenes. Annu Rev Neurosci. 1989;12:103–126. doi: 10.1146/annurev.ne.12.030189.000535. [DOI] [PubMed] [Google Scholar]
  41. Walsh F. S., Furness J., Moore S. E., Ashton S., Doherty P. Use of the neural cell adhesion molecule VASE exon by neurons is associated with a specific down-regulation of neural cell adhesion molecule-dependent neurite outgrowth in the developing cerebellum and hippocampus. J Neurochem. 1992 Nov;59(5):1959–1962. doi: 10.1111/j.1471-4159.1992.tb11033.x. [DOI] [PubMed] [Google Scholar]
  42. Wanaka A., Johnson E. M., Jr, Milbrandt J. Localization of FGF receptor mRNA in the adult rat central nervous system by in situ hybridization. Neuron. 1990 Sep;5(3):267–281. doi: 10.1016/0896-6273(90)90164-b. [DOI] [PubMed] [Google Scholar]
  43. Williams E. J., Doherty P., Turner G., Reid R. A., Hemperly J. J., Walsh F. S. Calcium influx into neurons can solely account for cell contact-dependent neurite outgrowth stimulated by transfected L1. J Cell Biol. 1992 Nov;119(4):883–892. doi: 10.1083/jcb.119.4.883. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES