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. 1993 Jul 1;122(1):181–189. doi: 10.1083/jcb.122.1.181

Thy-1 antibody-triggered neurite outgrowth requires an influx of calcium into neurons via N- and L-type calcium channels

PMCID: PMC2119600  PMID: 8100230

Abstract

We present evidence that the neurite out-growth stimulated by the binding of Thy-1 antibodies to PC12 cells is mediated by calcium influx through both N- and L-type calcium channels. PC12 cells cultured on a noncellular substratum in the presence of NGF, or on a cellular substratum in the absence of NGF, responded to soluble Thy-1 antibody by extending longer neurites. The response required bivalent antibody and could be blocked by removing Thy-1 from the surface of PC12 cells with phosphatidylinositol specific phospholipase C. The response could also be blocked by reducing extracellular calcium to 0.25 mM, or by antagonists of L- and N-type calcium channels. Additionally, the response could be fully inhibited by preloading PC12 cells with BAPTA/AM which buffers changes in intracellular calcium. A heterotrimeric G-protein is also implicated in the pathway as the response could be fully inhibited by pertussis toxin. These data suggest that antibody-induced clustering of Thy-1 stimulates neurite outgrowth by activating a second messenger pathway that has previously been shown to underlie cell adhesion molecule (NCAM, N-cadherin, and L1), but not integrin or NGF-dependent neurite outgrowth.

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Selected References

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  1. Bandtlow C. E., Schmidt M. F., Hassinger T. D., Schwab M. E., Kater S. B. Role of intracellular calcium in NI-35-evoked collapse of neuronal growth cones. Science. 1993 Jan 1;259(5091):80–83. doi: 10.1126/science.8418499. [DOI] [PubMed] [Google Scholar]
  2. Birren S. J., Anderson D. J. A v-myc-immortalized sympathoadrenal progenitor cell line in which neuronal differentiation is initiated by FGF but not NGF. Neuron. 1990 Feb;4(2):189–201. doi: 10.1016/0896-6273(90)90094-v. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. 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]
  6. 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]
  7. Doherty P., Walsh F. S. Control of Thy-1 glycoprotein expression in cultures of PC12 cells. J Neurochem. 1987 Aug;49(2):610–616. doi: 10.1111/j.1471-4159.1987.tb02907.x. [DOI] [PubMed] [Google Scholar]
  8. Gennarini G., Durbec P., Boned A., Rougon G., Goridis C. Transfected F3/F11 neuronal cell surface protein mediates intercellular adhesion and promotes neurite outgrowth. Neuron. 1991 Apr;6(4):595–606. doi: 10.1016/0896-6273(91)90062-5. [DOI] [PubMed] [Google Scholar]
  9. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  10. Gunter K. C., Germain R. N., Kroczek R. A., Saito T., Yokoyama W. M., Chan C., Weiss A., Shevach E. M. Thy-1-mediated T-cell activation requires co-expression of CD3/Ti complex. Nature. 1987 Apr 2;326(6112):505–507. doi: 10.1038/326505a0. [DOI] [PubMed] [Google Scholar]
  11. He H. T., Naquet P., Caillol D., Pierres M. Thy-1 supports adhesion of mouse thymocytes to thymic epithelial cells through a Ca2(+)-independent mechanism. J Exp Med. 1991 Feb 1;173(2):515–518. doi: 10.1084/jem.173.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Igarashi M., Strittmatter S. M., Vartanian T., Fishman M. C. Mediation by G proteins of signals that cause collapse of growth cones. Science. 1993 Jan 1;259(5091):77–79. doi: 10.1126/science.8418498. [DOI] [PubMed] [Google Scholar]
  13. Johnson P. W., Abramow-Newerly W., Seilheimer B., Sadoul R., Tropak M. B., Arquint M., Dunn R. J., Schachner M., Roder J. C. Recombinant myelin-associated glycoprotein confers neural adhesion and neurite outgrowth function. Neuron. 1989 Sep;3(3):377–385. doi: 10.1016/0896-6273(89)90262-6. [DOI] [PubMed] [Google Scholar]
  14. Kadmon G., Kowitz A., Altevogt P., Schachner M. The neural cell adhesion molecule N-CAM enhances L1-dependent cell-cell interactions. J Cell Biol. 1990 Jan;110(1):193–208. doi: 10.1083/jcb.110.1.193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kater S. B., Mills L. R. Regulation of growth cone behavior by calcium. J Neurosci. 1991 Apr;11(4):891–899. doi: 10.1523/JNEUROSCI.11-04-00891.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kuhn T. B., Stoeckli E. T., Condrau M. A., Rathjen F. G., Sonderegger P. Neurite outgrowth on immobilized axonin-1 is mediated by a heterophilic interaction with L1(G4). J Cell Biol. 1991 Nov;115(4):1113–1126. doi: 10.1083/jcb.115.4.1113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lipton S. A., Leifer D., Barnstable C. J. Selectivity of Thy-1 monoclonal antibodies in enhancing neurite outgrowth. Neurosci Lett. 1992 Mar 16;137(1):75–77. doi: 10.1016/0304-3940(92)90302-n. [DOI] [PubMed] [Google Scholar]
  18. Mahanthappa N. K., Patterson P. H. Thy-1 involvement in neurite outgrowth: perturbation by antibodies, phospholipase C, and mutation. Dev Biol. 1992 Mar;150(1):47–59. doi: 10.1016/0012-1606(92)90006-3. [DOI] [PubMed] [Google Scholar]
  19. Morris R. Thy-1 in developing nervous tissue. Dev Neurosci. 1985;7(3):133–160. doi: 10.1159/000112283. [DOI] [PubMed] [Google Scholar]
  20. Parekh R. B., Tse A. G., Dwek R. A., Williams A. F., Rademacher T. W. Tissue-specific N-glycosylation, site-specific oligosaccharide patterns and lentil lectin recognition of rat Thy-1. EMBO J. 1987 May;6(5):1233–1244. doi: 10.1002/j.1460-2075.1987.tb02359.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rasmussen H. Ernst Klenk lecture, November 1989. The complexities of intracellular Ca2+ signalling. Biol Chem Hoppe Seyler. 1990 Mar;371(3):191–206. [PubMed] [Google Scholar]
  22. Reber B. F., Reuter H. Dependence of cytosolic calcium in differentiating rat pheochromocytoma cells on calcium channels and intracellular stores. J Physiol. 1991 Apr;435:145–162. doi: 10.1113/jphysiol.1991.sp018502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Saleh M., Lang R. J., Bartlett P. F. Thy-1-mediated regulation of a low-threshold transient calcium current in cultured sensory neurons. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4543–4547. doi: 10.1073/pnas.85.12.4543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Stefanová I., Horejsí V., Ansotegui I. J., Knapp W., Stockinger H. GPI-anchored cell-surface molecules complexed to protein tyrosine kinases. Science. 1991 Nov 15;254(5034):1016–1019. doi: 10.1126/science.1719635. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Togari A., Dickens G., Kuzuya H., Guroff G. The effect of fibroblast growth factor on PC12 cells. J Neurosci. 1985 Feb;5(2):307–316. doi: 10.1523/JNEUROSCI.05-02-00307.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Volarević S., Burns C. M., Sussman J. J., Ashwell J. D. Intimate association of Thy-1 and the T-cell antigen receptor with the CD45 tyrosine phosphatase. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7085–7089. doi: 10.1073/pnas.87.18.7085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Xue G. P., Rivero B. P., Morris R. J. The surface glycoprotein Thy-1 is excluded from growing axons during development: a study of the expression of Thy-1 during axogenesis in hippocampus and hindbrain. Development. 1991 May;112(1):161–176. doi: 10.1242/dev.112.1.161. [DOI] [PubMed] [Google Scholar]

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