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. 1987 Mar 1;104(3):623–634. doi: 10.1083/jcb.104.3.623

Embryonic neural retinal cell response to extracellular matrix proteins: developmental changes and effects of the cell substratum attachment antibody (CSAT)

PMCID: PMC2114555  PMID: 3493247

Abstract

Cell attachment and neurite outgrowth by embryonic neural retinal cells were measured in separate quantitative assays to define differences in substrate preference and to demonstrate developmentally regulated changes in cellular response to different extracellular matrix glycoproteins. Cells attached to laminin, fibronectin, and collagen IV in a concentration-dependent fashion, though fibronectin was less effective for attachment than the other two substrates. Neurite outgrowth was much more extensive on laminin than on fibronectin or collagen IV. These results suggest that different substrates have distinct effects on neuronal differentiation. Neural retinal cell attachment and neurite outgrowth were inhibited on all three substrates by two antibodies, cell substratum attachment antibody (CSAT) and JG22, which recognize a cell surface glycoprotein complex required for cell interactions with several extracellular matrix constituents. In addition, retinal cells grew neurites on substrates coated with the CSAT antibodies. These results suggest that cell surface molecules recognized by this antibody are directly involved in cell attachment and neurite extension. Neural retinal cells from embryos of different ages varied in their capacity to interact with extracellular matrix substrates. Cells of all ages, embryonic day 6 (E6) to E12, attached to collagen IV and CSAT antibody substrates. In contrast, cell attachment to laminin and fibronectin diminished with increasing embryonic age. Age-dependent differences were found in the profile of proteins precipitated by the CSAT antibody, raising the possibility that modifications of these proteins are responsible for the dramatic changes in substrate preference of retinal cells between E6 and E12.

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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. Adler R., Manthorpe M., Skaper S. D., Varon S. Polyornithine-attached neurite-promoting factors (PNPFs). Culture sources and responsive neurons. Brain Res. 1981 Feb 9;206(1):129–144. doi: 10.1016/0006-8993(81)90105-0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Argraves W. S., Pytela R., Suzuki S., Millán J. L., Pierschbacher M. D., Ruoslahti E. cDNA sequences from the alpha subunit of the fibronectin receptor predict a transmembrane domain and a short cytoplasmic peptide. J Biol Chem. 1986 Oct 5;261(28):12922–12924. [PubMed] [Google Scholar]
  6. 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]
  7. Bottenstein J. E., Skaper S. D., Varon S. S., Sato G. H. Selective survival of neurons from chick embryo sensory ganglionic dissociates utilizing serum-free supplemented medium. Exp Cell Res. 1980 Jan;125(1):183–190. doi: 10.1016/0014-4827(80)90202-5. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Carbonetto S., Gruver M. M., Turner D. C. Nerve fiber growth in culture on fibronectin, collagen, and glycosaminoglycan substrates. J Neurosci. 1983 Nov;3(11):2324–2335. doi: 10.1523/JNEUROSCI.03-11-02324.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chapman A. E. Characterization of a 140Kd cell surface glycoprotein involved in myoblast adhesion. J Cell Biochem. 1984;25(2):109–121. doi: 10.1002/jcb.240250206. [DOI] [PubMed] [Google Scholar]
  11. Chen W. T., Greve J. M., Gottlieb D. I., Singer S. J. Immunocytochemical localization of 140 kD cell adhesion molecules in cultured chicken fibroblasts, and in chicken smooth muscle and intestinal epithelial tissues. J Histochem Cytochem. 1985 Jun;33(6):576–586. doi: 10.1177/33.6.3889142. [DOI] [PubMed] [Google Scholar]
  12. Chen W. T., Hasegawa E., Hasegawa T., Weinstock C., Yamada K. M. Development of cell surface linkage complexes in cultured fibroblasts. J Cell Biol. 1985 Apr;100(4):1103–1114. doi: 10.1083/jcb.100.4.1103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Collins F. Induction of neurite outgrowth by a conditioned-medium factor bound to the culture substratum. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5210–5213. doi: 10.1073/pnas.75.10.5210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Collins F., Lee M. R. Reversible developmental change in the ability of ciliary ganglion neurons to extend neurites in culture. J Neurosci. 1982 Apr;2(4):424–430. doi: 10.1523/JNEUROSCI.02-04-00424.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Coughlin M. D., Bloom E. M., Black I. B. Characterization of a neuronal growth factor from mouse heart-cell-conditioned medium. Dev Biol. 1981 Feb;82(1):56–68. doi: 10.1016/0012-1606(81)90428-0. [DOI] [PubMed] [Google Scholar]
  17. Damsky C. H., Knudsen K. A., Bradley D., Buck C. A., Horwitz A. F. Distribution of the cell substratum attachment (CSAT) antigen on myogenic and fibroblastic cells in culture. J Cell Biol. 1985 May;100(5):1528–1539. doi: 10.1083/jcb.100.5.1528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Davis G. E., Manthorpe M., Engvall E., Varon S. Isolation and characterization of rat schwannoma neurite-promoting factor: evidence that the factor contains laminin. J Neurosci. 1985 Oct;5(10):2662–2671. doi: 10.1523/JNEUROSCI.05-10-02662.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Decker C., Greggs R., Duggan K., Stubbs J., Horwitz A. Adhesive multiplicity in the interaction of embryonic fibroblasts and myoblasts with extracellular matrices. J Cell Biol. 1984 Oct;99(4 Pt 1):1398–1404. doi: 10.1083/jcb.99.4.1398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Duband J. L., Rocher S., Chen W. T., Yamada K. M., Thiery J. P. Cell adhesion and migration in the early vertebrate embryo: location and possible role of the putative fibronectin receptor complex. J Cell Biol. 1986 Jan;102(1):160–178. doi: 10.1083/jcb.102.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Faivre-Bauman A., Puymirat J., Loudes C., Barret A., Tixier-Vidal A. Laminin promotes attachment and neurite elongation of fetal hypothalamic neurons grown in serum-free medium. Neurosci Lett. 1984 Jan 27;44(1):83–89. doi: 10.1016/0304-3940(84)90225-8. [DOI] [PubMed] [Google Scholar]
  23. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  24. Goldberg S., Coulombre A. J. Topographical development of the ganglion cell fiber layer in the chick retina. A whole mount study. J Comp Neurol. 1972 Dec;146(4):507–518. doi: 10.1002/cne.901460406. [DOI] [PubMed] [Google Scholar]
  25. Greve J. M., Gottlieb D. I. Monoclonal antibodies which alter the morphology of cultured chick myogenic cells. J Cell Biochem. 1982;18(2):221–229. doi: 10.1002/jcb.1982.240180209. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. Halfter W., Deiss S., Schwarz U. The formation of the axonal pattern in the embryonic avian retina. J Comp Neurol. 1985 Feb 22;232(4):466–480. doi: 10.1002/cne.902320405. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Hasegawa T., Hasegawa E., Chen W. T., Yamada K. M. Characterization of a membrane-associated glycoprotein complex implicated in cell adhesion to fibronectin. J Cell Biochem. 1985;28(4):307–318. doi: 10.1002/jcb.240280409. [DOI] [PubMed] [Google Scholar]
  31. Horwitz A., Duggan K., Buck C., Beckerle M. C., Burridge K. Interaction of plasma membrane fibronectin receptor with talin--a transmembrane linkage. Nature. 1986 Apr 10;320(6062):531–533. doi: 10.1038/320531a0. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Kahn A. J. Ganglion cell formation in the chick neural retina. Brain Res. 1973 Dec 7;63:285–290. doi: 10.1016/0006-8993(73)90095-4. [DOI] [PubMed] [Google Scholar]
  34. Knudsen K. A., Horwitz A. F., Buck C. A. A monoclonal antibody identifies a glycoprotein complex involved in cell-substratum adhesion. Exp Cell Res. 1985 Mar;157(1):218–226. doi: 10.1016/0014-4827(85)90164-8. [DOI] [PubMed] [Google Scholar]
  35. Koda J. E., Rapraeger A., Bernfield M. Heparan sulfate proteoglycans from mouse mammary epithelial cells. Cell surface proteoglycan as a receptor for interstitial collagens. J Biol Chem. 1985 Jul 5;260(13):8157–8162. [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. Lander A. D., Fujii D. K., Gospodarowicz D., Reichardt L. F. Characterization of a factor that promotes neurite outgrowth: evidence linking activity to a heparan sulfate proteoglycan. J Cell Biol. 1982 Sep;94(3):574–585. doi: 10.1083/jcb.94.3.574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Lander A. D., Fujii D. K., Reichardt L. F. Purification of a factor that promotes neurite outgrowth: isolation of laminin and associated molecules. J Cell Biol. 1985 Sep;101(3):898–913. doi: 10.1083/jcb.101.3.898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lander A. D., Tomaselli K., Calof A. L., Reichardt L. F. Studies on extracellular matrix components that promote neurite outgrowth. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 2):611–623. doi: 10.1101/sqb.1983.048.01.065. [DOI] [PubMed] [Google Scholar]
  40. Leptin M. The fibronectin receptor family. Nature. 1986 Jun 19;321(6072):728–728. doi: 10.1038/321728a0. [DOI] [PubMed] [Google Scholar]
  41. Letourneau P. C. Cell-to-substratum adhesion and guidance of axonal elongation. Dev Biol. 1975 May;44(1):92–101. doi: 10.1016/0012-1606(75)90379-6. [DOI] [PubMed] [Google Scholar]
  42. MacLeish P. R., Barnstable C. J., Townes-Anderson E. Use of a monoclonal antibody as a substrate for mature neurons in vitro. Proc Natl Acad Sci U S A. 1983 Nov;80(22):7014–7018. doi: 10.1073/pnas.80.22.7014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]
  44. Manthorpe M., Fagnani R., Skaper S. D., Varon S. An automated colorimetric microassay for neuronotrophic factors. Brain Res. 1986 Mar;390(2):191–198. doi: 10.1016/s0006-8993(86)80227-x. [DOI] [PubMed] [Google Scholar]
  45. 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]
  46. Pytela R., Pierschbacher M. D., Ginsberg M. H., Plow E. F., Ruoslahti E. Platelet membrane glycoprotein IIb/IIIa: member of a family of Arg-Gly-Asp--specific adhesion receptors. Science. 1986 Mar 28;231(4745):1559–1562. doi: 10.1126/science.2420006. [DOI] [PubMed] [Google Scholar]
  47. Pytela R., Pierschbacher M. D., Ruoslahti E. Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor. Cell. 1985 Jan;40(1):191–198. doi: 10.1016/0092-8674(85)90322-8. [DOI] [PubMed] [Google Scholar]
  48. Rager G. H. Development of the retinotectal projection in the chicken. Adv Anat Embryol Cell Biol. 1980;63:I-VIII, 1-90. [PubMed] [Google Scholar]
  49. Raper J. A., Bastiani M., Goodman C. S. Pathfinding by neuronal growth cones in grasshopper embryos. I. Divergent choices made by the growth cones of sibling neurons. J Neurosci. 1983 Jan;3(1):20–30. doi: 10.1523/JNEUROSCI.03-01-00020.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Rein A., Rubin H. Effects of local cell concentrations upon the growth of chick embryo cells in tissue culture. Exp Cell Res. 1968 Mar;49(3):666–678. doi: 10.1016/0014-4827(68)90213-9. [DOI] [PubMed] [Google Scholar]
  51. Rogers S. L., Edson K. J., Letourneau P. C., McLoon S. C. Distribution of laminin in the developing peripheral nervous system of the chick. Dev Biol. 1986 Feb;113(2):429–435. doi: 10.1016/0012-1606(86)90177-6. [DOI] [PubMed] [Google Scholar]
  52. 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]
  53. Ruoslahti E., Pierschbacher M. D. Arg-Gly-Asp: a versatile cell recognition signal. Cell. 1986 Feb 28;44(4):517–518. doi: 10.1016/0092-8674(86)90259-x. [DOI] [PubMed] [Google Scholar]
  54. Smalheiser N. R., Crain S. M., Reid L. M. Laminin as a substrate for retinal axons in vitro. Brain Res. 1984 Jan;314(1):136–140. doi: 10.1016/0165-3806(84)90184-6. [DOI] [PubMed] [Google Scholar]
  55. Tamkun J. W., DeSimone D. W., Fonda D., Patel R. S., Buck C., Horwitz A. F., Hynes R. O. Structure of integrin, a glycoprotein involved in the transmembrane linkage between fibronectin and actin. Cell. 1986 Jul 18;46(2):271–282. doi: 10.1016/0092-8674(86)90744-0. [DOI] [PubMed] [Google Scholar]
  56. Taylor J. S., Roberts A. The early development of the primary sensory neurones in an amphibian embryo: a scanning electron microscope study. J Embryol Exp Morphol. 1983 Jun;75:49–66. [PubMed] [Google Scholar]
  57. Thanos S., Bonhoeffer F. Investigations on the development and topographic order of retinotectal axons: anterograde and retrograde staining of axons and perikarya with rhodamine in vivo. J Comp Neurol. 1983 Oct 1;219(4):420–430. doi: 10.1002/cne.902190404. [DOI] [PubMed] [Google Scholar]
  58. 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]
  59. 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]
  60. Unsicker K., Skaper S. D., Varon S. Developmental changes in the responses of rat chromaffin cells to neuronotrophic and neurite-promoting factors. Dev Biol. 1985 Oct;111(2):425–433. doi: 10.1016/0012-1606(85)90495-6. [DOI] [PubMed] [Google Scholar]
  61. Vlodavsky I., Levi A., Lax I., Fuks Z., Schlessinger J. Induction of cell attachment and morphological differentiation in a pheochromocytoma cell line and embryonal sensory cells by the extracellular matrix. Dev Biol. 1982 Oct;93(2):285–300. doi: 10.1016/0012-1606(82)90118-x. [DOI] [PubMed] [Google Scholar]
  62. Zhu B. C., Fisher S. F., Pande H., Calaycay J., Shively J. E., Laine R. A. Human placental (fetal) fibronectin: increased glycosylation and higher protease resistance than plasma fibronectin. Presence of polylactosamine glycopeptides and properties of a 44-kilodalton chymotryptic collagen-binding domain: difference from human plasma fibronectin. J Biol Chem. 1984 Mar 25;259(6):3962–3970. [PubMed] [Google Scholar]

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