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. 1988 Sep 1;107(3):1177–1187. doi: 10.1083/jcb.107.3.1177

N-cadherin, NCAM, and integrins promote retinal neurite outgrowth on astrocytes in vitro

PMCID: PMC2115273  PMID: 3262111

Abstract

Retinal ganglion neurons extend axons that grow along astroglial cell surfaces in the developing optic pathway. To identify the molecules that may mediate axon extension in vivo, antibodies to neuronal cell surface proteins were tested for their effects on neurite outgrowth by embryonic chick retinal neurons cultured on astrocyte monolayers. Neurite outgrowth by retinal neurons from embryonic day 7 (E7) and E11 chick embryos depended on the function of a calcium-dependent cell adhesion molecule (N-cadherin) and beta 1-class integrin extracellular matrix receptors. The inhibitory effects of either antibody on process extension could not be accounted for by a reduction in the attachment of neurons to astrocytes. The role of a third cell adhesion molecule, NCAM, changed during development. Anti-NCAM had no detectable inhibitory effects on neurite outgrowth by E7 retinal neurons. In contrast, E11 retinal neurite outgrowth was strongly dependent on NCAM function. Thus, N-cadherin, integrins, and NCAM are likely to regulate axon extension in the optic pathway, and their relative importance varies with developmental age.

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Selected References

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  1. Adler R., Jerdan J., Hewitt A. T. Responses of cultured neural retinal cells to substratum-bound laminin and other extracellular matrix molecules. Dev Biol. 1985 Nov;112(1):100–114. doi: 10.1016/0012-1606(85)90124-1. [DOI] [PubMed] [Google Scholar]
  2. Akers R. M., Mosher D. F., Lilien J. E. Promotion of retinal neurite outgrowth by substratum-bound fibronectin. Dev Biol. 1981 Aug;86(1):179–188. doi: 10.1016/0012-1606(81)90328-6. [DOI] [PubMed] [Google Scholar]
  3. Akiyama S. K., Yamada S. S., Yamada K. M. Characterization of a 140-kD avian cell surface antigen as a fibronectin-binding molecule. J Cell Biol. 1986 Feb;102(2):442–448. doi: 10.1083/jcb.102.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Bixby J. L., Lilien J., Reichardt L. F. Identification of the major proteins that promote neuronal process outgrowth on Schwann cells in vitro. J Cell Biol. 1988 Jul;107(1):353–361. doi: 10.1083/jcb.107.1.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Bixby J. L., Reichardt L. F. Effects of antibodies to neural cell adhesion molecule (N-CAM) on the differentiation of neuromuscular contacts between ciliary ganglion neurons and myotubes in vitro. Dev Biol. 1987 Feb;119(2):363–372. doi: 10.1016/0012-1606(87)90041-8. [DOI] [PubMed] [Google Scholar]
  8. Bohn R. C., Reier P. J., Sourbeer E. B. Axonal interactions with connective tissue and glial substrata during optic nerve regeneration in Xenopus larvae and adults. Am J Anat. 1982 Dec;165(4):397–419. doi: 10.1002/aja.1001650405. [DOI] [PubMed] [Google Scholar]
  9. Bork T., Schabtach E., Grant P. Factors guiding optic fibers in developing Xenopus retina. J Comp Neurol. 1987 Oct 8;264(2):147–158. doi: 10.1002/cne.902640202. [DOI] [PubMed] [Google Scholar]
  10. Bovolenta P., Mason C. Growth cone morphology varies with position in the developing mouse visual pathway from retina to first targets. J Neurosci. 1987 May;7(5):1447–1460. doi: 10.1523/JNEUROSCI.07-05-01447.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Bozyczko D., Horwitz A. F. The participation of a putative cell surface receptor for laminin and fibronectin in peripheral neurite extension. J Neurosci. 1986 May;6(5):1241–1251. doi: 10.1523/JNEUROSCI.06-05-01241.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Buck C. A., Shea E., Duggan K., Horwitz A. F. Integrin (the CSAT antigen): functionality requires oligomeric integrity. J Cell Biol. 1986 Dec;103(6 Pt 1):2421–2428. doi: 10.1083/jcb.103.6.2421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chang S., Rathjen F. G., Raper J. A. Extension of neurites on axons is impaired by antibodies against specific neural cell surface glycoproteins. J Cell Biol. 1987 Feb;104(2):355–362. doi: 10.1083/jcb.104.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Cohen J., Burne J. F., McKinlay C., Winter J. The role of laminin and the laminin/fibronectin receptor complex in the outgrowth of retinal ganglion cell axons. Dev Biol. 1987 Aug;122(2):407–418. doi: 10.1016/0012-1606(87)90305-8. [DOI] [PubMed] [Google Scholar]
  15. Cohen J., Burne J. F., Winter J., Bartlett P. Retinal ganglion cells lose response to laminin with maturation. 1986 Jul 31-Aug 6Nature. 322(6078):465–467. doi: 10.1038/322465a0. [DOI] [PubMed] [Google Scholar]
  16. Crittenden S. L., Pratt R. S., Cook J. H., Balsamo J., Lilien J. Immunologically unique and common domains within a family of proteins related to the retina Ca2+-dependent cell adhesion molecule, NcalCAM. Development. 1987 Dec;101(4):729–740. doi: 10.1242/dev.101.4.729. [DOI] [PubMed] [Google Scholar]
  17. Crossin K. L., Hoffman S., Grumet M., Thiery J. P., Edelman G. M. Site-restricted expression of cytotactin during development of the chicken embryo. J Cell Biol. 1986 May;102(5):1917–1930. doi: 10.1083/jcb.102.5.1917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Easter S. S., Jr, Bratton B., Scherer S. S. Growth-related order of the retinal fiber layer in goldfish. J Neurosci. 1984 Aug;4(8):2173–2190. doi: 10.1523/JNEUROSCI.04-08-02173.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Eisenbarth G. S., Walsh F. S., Nirenberg M. Monoclonal antibody to a plasma membrane antigen of neurons. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4913–4917. doi: 10.1073/pnas.76.10.4913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Fallon J. R. Preferential outgrowth of central nervous system neurites on astrocytes and Schwann cells as compared with nonglial cells in vitro. J Cell Biol. 1985 Jan;100(1):198–207. doi: 10.1083/jcb.100.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Fraser S. E., Murray B. A., Chuong C. M., Edelman G. M. Alteration of the retinotectal map in Xenopus by antibodies to neural cell adhesion molecules. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4222–4226. doi: 10.1073/pnas.81.13.4222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Grumet M., Edelman G. M. Neuron-glia cell adhesion molecule interacts with neurons and astroglia via different binding mechanisms. J Cell Biol. 1988 Feb;106(2):487–503. doi: 10.1083/jcb.106.2.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Grunwald G. B., Pratt R. S., Lilien J. Enzymic dissection of embryonic cell adhesive mechanisms. III. Immunological identification of a component of the calcium-dependent adhesive system of embryonic chick neural retina cells. J Cell Sci. 1982 Jun;55:69–83. doi: 10.1242/jcs.55.1.69. [DOI] [PubMed] [Google Scholar]
  25. Halfter W., Deiss S. Axon growth in embryonic chick and quail retinal whole mounts in vitro. Dev Biol. 1984 Apr;102(2):344–355. doi: 10.1016/0012-1606(84)90199-4. [DOI] [PubMed] [Google Scholar]
  26. Halfter W., Deiss S. Axonal pathfinding in organ-cultured embryonic avian retinae. Dev Biol. 1986 Apr;114(2):296–310. doi: 10.1016/0012-1606(86)90194-6. [DOI] [PubMed] [Google Scholar]
  27. Halfter W., Newgreen D. F., Sauter J., Schwarz U. Oriented axon outgrowth from avian embryonic retinae in culture. Dev Biol. 1983 Jan;95(1):56–64. doi: 10.1016/0012-1606(83)90006-4. [DOI] [PubMed] [Google Scholar]
  28. Halfter W., Reckhaus W., Kröger S. Nondirected axonal growth on basal lamina from avian embryonic neural retina. J Neurosci. 1987 Nov;7(11):3712–3722. doi: 10.1523/JNEUROSCI.07-11-03712.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Hall D. E., Neugebauer K. M., Reichardt L. F. Embryonic neural retinal cell response to extracellular matrix proteins: developmental changes and effects of the cell substratum attachment antibody (CSAT). J Cell Biol. 1987 Mar;104(3):623–634. doi: 10.1083/jcb.104.3.623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. Hatta K., Takeichi M. Expression of N-cadherin adhesion molecules associated with early morphogenetic events in chick development. Nature. 1986 Apr 3;320(6061):447–449. doi: 10.1038/320447a0. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Horwitz A., Duggan K., Greggs R., Decker C., Buck C. The cell substrate attachment (CSAT) antigen has properties of a receptor for laminin and fibronectin. J Cell Biol. 1985 Dec;101(6):2134–2144. doi: 10.1083/jcb.101.6.2134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Hynes R. O. Integrins: a family of cell surface receptors. Cell. 1987 Feb 27;48(4):549–554. doi: 10.1016/0092-8674(87)90233-9. [DOI] [PubMed] [Google Scholar]
  36. Kahn A. J. An autoradiographic analysis of the time of appearance of neurons in the developing chick neural retina. Dev Biol. 1974 May;38(1):30–40. doi: 10.1016/0012-1606(74)90256-5. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Lagenaur C., Lemmon V. An L1-like molecule, the 8D9 antigen, is a potent substrate for neurite extension. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7753–7757. doi: 10.1073/pnas.84.21.7753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lemmon V. Localization of a filamin-like protein in glia of the chick central nervous system. J Neurosci. 1986 Jan;6(1):43–51. doi: 10.1523/JNEUROSCI.06-01-00043.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. 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]
  41. Lemmon V. Monoclonal antibodies specific for glia in the chick nervous system. Brain Res. 1985 Nov;355(1):111–120. doi: 10.1016/0165-3806(85)90010-0. [DOI] [PubMed] [Google Scholar]
  42. Lemmon V., Staros E. B., Perry H. E., Gottlieb D. I. A monoclonal antibody which binds to the surface of chick brain cells and myotubes: cell selectivity and properties of the antigen. Brain Res. 1982 Mar;255(3):349–360. doi: 10.1016/0165-3806(82)90003-7. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. McCaffery C. A., Raju T. R., Bennett M. R. Effects of cultured astroglia on the survival of neonatal rat retinal ganglion cells in vitro. Dev Biol. 1984 Aug;104(2):441–448. doi: 10.1016/0012-1606(84)90100-3. [DOI] [PubMed] [Google Scholar]
  45. Nagafuchi A., Shirayoshi Y., Okazaki K., Yasuda K., Takeichi M. Transformation of cell adhesion properties by exogenously introduced E-cadherin cDNA. Nature. 1987 Sep 24;329(6137):341–343. doi: 10.1038/329341a0. [DOI] [PubMed] [Google Scholar]
  46. Neff N. T., Lowrey C., Decker C., Tovar A., Damsky C., Buck C., Horwitz A. F. A monoclonal antibody detaches embryonic skeletal muscle from extracellular matrices. J Cell Biol. 1982 Nov;95(2 Pt 1):654–666. doi: 10.1083/jcb.95.2.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. 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]
  48. Noble M., Albrechtsen M., Møller C., Lyles J., Bock E., Goridis C., Watanabe M., Rutishauser U. Glial cells express N-CAM/D2-CAM-like polypeptides in vitro. Nature. 1985 Aug 22;316(6030):725–728. doi: 10.1038/316725a0. [DOI] [PubMed] [Google Scholar]
  49. Noble M., Fok-Seang J., Cohen J. Glia are a unique substrate for the in vitro growth of central nervous system neurons. J Neurosci. 1984 Jul;4(7):1892–1903. doi: 10.1523/JNEUROSCI.04-07-01892.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Price J., Hynes R. O. Astrocytes in culture synthesize and secrete a variant form of fibronectin. J Neurosci. 1985 Aug;5(8):2205–2211. doi: 10.1523/JNEUROSCI.05-08-02205.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Raff M. C., Abney E. R., Cohen J., Lindsay R., Noble M. Two types of astrocytes in cultures of developing rat white matter: differences in morphology, surface gangliosides, and growth characteristics. J Neurosci. 1983 Jun;3(6):1289–1300. doi: 10.1523/JNEUROSCI.03-06-01289.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Ruoslahti E., Pierschbacher M. D. New perspectives in cell adhesion: RGD and integrins. Science. 1987 Oct 23;238(4826):491–497. doi: 10.1126/science.2821619. [DOI] [PubMed] [Google Scholar]
  53. Rutishauser U., Grumet M., Edelman G. M. Neural cell adhesion molecule mediates initial interactions between spinal cord neurons and muscle cells in culture. J Cell Biol. 1983 Jul;97(1):145–152. doi: 10.1083/jcb.97.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. SPERRY R. W. CHEMOAFFINITY IN THE ORDERLY GROWTH OF NERVE FIBER PATTERNS AND CONNECTIONS. Proc Natl Acad Sci U S A. 1963 Oct;50:703–710. doi: 10.1073/pnas.50.4.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Schlosshauer B., Schwarz U., Rutishauser U. Topological distribution of different forms of neural cell adhesion molecule in the developing chick visual system. Nature. 1984 Jul 12;310(5973):141–143. doi: 10.1038/310141a0. [DOI] [PubMed] [Google Scholar]
  56. 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]
  57. Smith G. M., Miller R. H., Silver J. Changing role of forebrain astrocytes during development, regenerative failure, and induced regeneration upon transplantation. J Comp Neurol. 1986 Sep 1;251(1):23–43. doi: 10.1002/cne.902510103. [DOI] [PubMed] [Google Scholar]
  58. Thanos S., Bonhoeffer F., Rutishauser U. Fiber-fiber interaction and tectal cues influence the development of the chicken retinotectal projection. Proc Natl Acad Sci U S A. 1984 Mar;81(6):1906–1910. doi: 10.1073/pnas.81.6.1906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Thor G., Pollerberg E. G., Schachner M. Molecular association of two neural cell adhesion molecules within the surface membrane of cultured mouse neuroblastoma cells. Neurosci Lett. 1986 May 15;66(2):121–126. doi: 10.1016/0304-3940(86)90176-x. [DOI] [PubMed] [Google Scholar]
  60. Timpl R., Rohde H., Robey P. G., Rennard S. I., Foidart J. M., Martin G. R. Laminin--a glycoprotein from basement membranes. J Biol Chem. 1979 Oct 10;254(19):9933–9937. [PubMed] [Google Scholar]
  61. Tomaselli K. J., Damsky C. H., Reichardt L. F. Interactions of a neuronal cell line (PC12) with laminin, collagen IV, and fibronectin: identification of integrin-related glycoproteins involved in attachment and process outgrowth. J Cell Biol. 1987 Nov;105(5):2347–2358. doi: 10.1083/jcb.105.5.2347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Tomaselli K. J., Neugebauer K. M., Bixby J. L., Lilien J., Reichardt L. F. N-cadherin and integrins: two receptor systems that mediate neuronal process outgrowth on astrocyte surfaces. Neuron. 1988 Mar;1(1):33–43. doi: 10.1016/0896-6273(88)90207-3. [DOI] [PubMed] [Google Scholar]
  63. Tomaselli K. J., Reichardt L. F., Bixby J. L. Distinct molecular interactions mediate neuronal process outgrowth on non-neuronal cell surfaces and extracellular matrices. J Cell Biol. 1986 Dec;103(6 Pt 2):2659–2672. doi: 10.1083/jcb.103.6.2659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Watanabe M., Frelinger A. L., 3rd, Rutishauser U. Topography of N-CAM structural and functional determinants. I. Classification of monoclonal antibody epitopes. J Cell Biol. 1986 Nov;103(5):1721–1727. doi: 10.1083/jcb.103.5.1721. [DOI] [PMC free article] [PubMed] [Google Scholar]

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