Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1993 Jun 1;90(11):5327–5331. doi: 10.1073/pnas.90.11.5327

GAP-43 augments G protein-coupled receptor transduction in Xenopus laevis oocytes.

S M Strittmatter 1, S C Cannon 1, E M Ross 1, T Higashijima 1, M C Fishman 1
PMCID: PMC46709  PMID: 7685122

Abstract

The neuronal protein GAP-43 is thought to play a role in determining growth-cone motility, perhaps as an intracellular regulator of signal transduction, but its molecular mechanism of action has remained unclear. We find that GAP-43, when microinjected into Xenopus laevis oocytes, increases the oocyte response to G protein-coupled receptor agonists by 10- to 100-fold. Higher levels of GAP-43 cause a transient current flow, even without receptor stimulation. The GAP-43-induced current, like receptor-stimulated currents, is mediated by a calcium-activated chloride channel and can be desensitized by injection of inositol 1,4,5-trisphosphate. This suggests that neuronal GAP-43 may serve as an intracellular signal to greatly enhance the sensitivity of G protein-coupled receptor transduction.

Full text

PDF
5327

Images in this article

5328Fig. 1
on p.5328
5328Fig. 2
on p.5328

Selected References

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

  1. Alexander K. A., Wakim B. T., Doyle G. S., Walsh K. A., Storm D. R. Identification and characterization of the calmodulin-binding domain of neuromodulin, a neurospecific calmodulin-binding protein. J Biol Chem. 1988 Jun 5;263(16):7544–7549. [PubMed] [Google Scholar]
  2. Baetge E. E., Hammang J. P. Neurite outgrowth in PC12 cells deficient in GAP-43. Neuron. 1991 Jan;6(1):21–30. doi: 10.1016/0896-6273(91)90118-j. [DOI] [PubMed] [Google Scholar]
  3. Bauer P. H., Müller S., Puzicha M., Pippig S., Obermaier B., Helmreich E. J., Lohse M. J. Phosducin is a protein kinase A-regulated G-protein regulator. Nature. 1992 Jul 2;358(6381):73–76. doi: 10.1038/358073a0. [DOI] [PubMed] [Google Scholar]
  4. Benowitz L. I., Apostolides P. J., Perrone-Bizzozero N., Finklestein S. P., Zwiers H. Anatomical distribution of the growth-associated protein GAP-43/B-50 in the adult rat brain. J Neurosci. 1988 Jan;8(1):339–352. doi: 10.1523/JNEUROSCI.08-01-00339.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chapman E. R., Au D., Alexander K. A., Nicolson T. A., Storm D. R. Characterization of the calmodulin binding domain of neuromodulin. Functional significance of serine 41 and phenylalanine 42. J Biol Chem. 1991 Jan 5;266(1):207–213. [PubMed] [Google Scholar]
  6. Coggins P. J., Zwiers H. Evidence for a single protein kinase C-mediated phosphorylation site in rat brain protein B-50. J Neurochem. 1989 Dec;53(6):1895–1901. doi: 10.1111/j.1471-4159.1989.tb09259.x. [DOI] [PubMed] [Google Scholar]
  7. Cox E. C., Müller B., Bonhoeffer F. Axonal guidance in the chick visual system: posterior tectal membranes induce collapse of growth cones from the temporal retina. Neuron. 1990 Jan;4(1):31–37. doi: 10.1016/0896-6273(90)90441-h. [DOI] [PubMed] [Google Scholar]
  8. Davies J. A., Cook G. M., Stern C. D., Keynes R. J. Isolation from chick somites of a glycoprotein fraction that causes collapse of dorsal root ganglion growth cones. Neuron. 1990 Jan;4(1):11–20. doi: 10.1016/0896-6273(90)90439-m. [DOI] [PubMed] [Google Scholar]
  9. Dekker L. V., De Graan P. N., Oestreicher A. B., Versteeg D. H., Gispen W. H. Inhibition of noradrenaline release by antibodies to B-50 (GAP-43). Nature. 1989 Nov 2;342(6245):74–76. doi: 10.1038/342074a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Gianotti C., Nunzi M. G., Gispen W. H., Corradetti R. Phosphorylation of the presynaptic protein B-50 (GAP-43) is increased during electrically induced long-term potentiation. Neuron. 1992 May;8(5):843–848. doi: 10.1016/0896-6273(92)90198-m. [DOI] [PubMed] [Google Scholar]
  12. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  13. Goh J. W., Pennefather P. S. A pertussis toxin-sensitive G protein in hippocampal long-term potentiation. Science. 1989 May 26;244(4907):980–983. doi: 10.1126/science.2543072. [DOI] [PubMed] [Google Scholar]
  14. Haydon P. G., Man-Son-Hing H., Doyle R. T., Zoran M. FMRFamide modulation of secretory machinery underlying presynaptic inhibition of synaptic transmission requires a pertussis toxin-sensitive G-protein. J Neurosci. 1991 Dec;11(12):3851–3860. doi: 10.1523/JNEUROSCI.11-12-03851.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Haydon P. G., McCobb D. P., Kater S. B. Serotonin selectively inhibits growth cone motility and synaptogenesis of specific identified neurons. Science. 1984 Nov 2;226(4674):561–564. doi: 10.1126/science.6093252. [DOI] [PubMed] [Google Scholar]
  16. Higashijima T., Burnier J., Ross E. M. Regulation of Gi and Go by mastoparan, related amphiphilic peptides, and hydrophobic amines. Mechanism and structural determinants of activity. J Biol Chem. 1990 Aug 25;265(24):14176–14186. [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. Kusano K., Miledi R., Stinnakre J. Cholinergic and catecholaminergic receptors in the Xenopus oocyte membrane. J Physiol. 1982 Jul;328:143–170. doi: 10.1113/jphysiol.1982.sp014257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lankford K. L., DeMello F. G., Klein W. L. D1-type dopamine receptors inhibit growth cone motility in cultured retina neurons: evidence that neurotransmitters act as morphogenic growth regulators in the developing central nervous system. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2839–2843. doi: 10.1073/pnas.85.8.2839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lechleiter J., Girard S., Peralta E., Clapham D. Spiral calcium wave propagation and annihilation in Xenopus laevis oocytes. Science. 1991 Apr 5;252(5002):123–126. doi: 10.1126/science.2011747. [DOI] [PubMed] [Google Scholar]
  22. Lechleiter J., Hellmiss R., Duerson K., Ennulat D., David N., Clapham D., Peralta E. Distinct sequence elements control the specificity of G protein activation by muscarinic acetylcholine receptor subtypes. EMBO J. 1990 Dec;9(13):4381–4390. doi: 10.1002/j.1460-2075.1990.tb07888.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lockerbie R. O. The neuronal growth cone: a review of its locomotory, navigational and target recognition capabilities. Neuroscience. 1987 Mar;20(3):719–729. doi: 10.1016/0306-4522(87)90235-1. [DOI] [PubMed] [Google Scholar]
  24. Mattson M. P., Dou P., Kater S. B. Outgrowth-regulating actions of glutamate in isolated hippocampal pyramidal neurons. J Neurosci. 1988 Jun;8(6):2087–2100. doi: 10.1523/JNEUROSCI.08-06-02087.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Meiri K. F., Gordon-Weeks P. R. GAP-43 in growth cones is associated with areas of membrane that are tightly bound to substrate and is a component of a membrane skeleton subcellular fraction. J Neurosci. 1990 Jan;10(1):256–266. doi: 10.1523/JNEUROSCI.10-01-00256.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Meiri K. F., Pfenninger K. H., Willard M. B. Growth-associated protein, GAP-43, a polypeptide that is induced when neurons extend axons, is a component of growth cones and corresponds to pp46, a major polypeptide of a subcellular fraction enriched in growth cones. Proc Natl Acad Sci U S A. 1986 May;83(10):3537–3541. doi: 10.1073/pnas.83.10.3537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Moriarty T. M., Padrell E., Carty D. J., Omri G., Landau E. M., Iyengar R. Go protein as signal transducer in the pertussis toxin-sensitive phosphatidylinositol pathway. Nature. 1990 Jan 4;343(6253):79–82. doi: 10.1038/343079a0. [DOI] [PubMed] [Google Scholar]
  28. Oestreicher A. B., Van Dongen C. J., Zwiers H., Gispen W. H. Affinity-purified anti-B-50 protein antibody: interference with the function of the phosphoprotein B-50 in synaptic plasma membranes. J Neurochem. 1983 Aug;41(2):331–340. doi: 10.1111/j.1471-4159.1983.tb04747.x. [DOI] [PubMed] [Google Scholar]
  29. Parker E. M., Kameyama K., Higashijima T., Ross E. M. Reconstitutively active G protein-coupled receptors purified from baculovirus-infected insect cells. J Biol Chem. 1991 Jan 5;266(1):519–527. [PubMed] [Google Scholar]
  30. Parker I., Miledi R. Nonlinearity and facilitation in phosphoinositide signaling studied by the use of caged inositol trisphosphate in Xenopus oocytes. J Neurosci. 1989 Nov;9(11):4068–4077. doi: 10.1523/JNEUROSCI.09-11-04068.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Patterson P. H. On the importance of being inhibited, or saying no to growth cones. Neuron. 1988 Jun;1(4):263–267. doi: 10.1016/0896-6273(88)90074-8. [DOI] [PubMed] [Google Scholar]
  32. Raper J. A., Kapfhammer J. P. The enrichment of a neuronal growth cone collapsing activity from embryonic chick brain. Neuron. 1990 Jan;4(1):21–29. doi: 10.1016/0896-6273(90)90440-q. [DOI] [PubMed] [Google Scholar]
  33. Rodrigues P. dos S., Dowling J. E. Dopamine induces neurite retraction in retinal horizontal cells via diacylglycerol and protein kinase C. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9693–9697. doi: 10.1073/pnas.87.24.9693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schuch U., Lohse M. J., Schachner M. Neural cell adhesion molecules influence second messenger systems. Neuron. 1989 Jul;3(1):13–20. doi: 10.1016/0896-6273(89)90111-6. [DOI] [PubMed] [Google Scholar]
  35. Shea T. B., Perrone-Bizzozero N. I., Beermann M. L., Benowitz L. I. Phospholipid-mediated delivery of anti-GAP-43 antibodies into neuroblastoma cells prevents neuritogenesis. J Neurosci. 1991 Jun;11(6):1685–1690. doi: 10.1523/JNEUROSCI.11-06-01685.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Singer D., Boton R., Moran O., Dascal N. Short- and long-term desensitization of serotonergic response in Xenopus oocytes injected with brain RNA: roles for inositol 1,4,5-trisphosphate and protein kinase C. Pflugers Arch. 1990 Apr;416(1-2):7–16. doi: 10.1007/BF00370215. [DOI] [PubMed] [Google Scholar]
  37. Skene J. H. Axonal growth-associated proteins. Annu Rev Neurosci. 1989;12:127–156. doi: 10.1146/annurev.ne.12.030189.001015. [DOI] [PubMed] [Google Scholar]
  38. Skene J. H., Jacobson R. D., Snipes G. J., McGuire C. B., Norden J. J., Freeman J. A. A protein induced during nerve growth (GAP-43) is a major component of growth-cone membranes. Science. 1986 Aug 15;233(4765):783–786. doi: 10.1126/science.3738509. [DOI] [PubMed] [Google Scholar]
  39. Strittmatter S. M., Fishman M. C. The neuronal growth cone as a specialized transduction system. Bioessays. 1991 Mar;13(3):127–134. doi: 10.1002/bies.950130306. [DOI] [PubMed] [Google Scholar]
  40. Strittmatter S. M. GAP-43 as a modulator of G protein transduction in the growth cone. Perspect Dev Neurobiol. 1992;1(1):13–19. [PubMed] [Google Scholar]
  41. Strittmatter S. M., Valenzuela D., Kennedy T. E., Neer E. J., Fishman M. C. G0 is a major growth cone protein subject to regulation by GAP-43. Nature. 1990 Apr 26;344(6269):836–841. doi: 10.1038/344836a0. [DOI] [PubMed] [Google Scholar]
  42. Strittmatter S. M., Valenzuela D., Sudo Y., Linder M. E., Fishman M. C. An intracellular guanine nucleotide release protein for G0. GAP-43 stimulates isolated alpha subunits by a novel mechanism. J Biol Chem. 1991 Nov 25;266(33):22465–22471. [PubMed] [Google Scholar]
  43. Strittmatter S. M., Vartanian T., Fishman M. C. GAP-43 as a plasticity protein in neuronal form and repair. J Neurobiol. 1992 Jul;23(5):507–520. doi: 10.1002/neu.480230506. [DOI] [PubMed] [Google Scholar]
  44. Sudo Y., Valenzuela D., Beck-Sickinger A. G., Fishman M. C., Strittmatter S. M. Palmitoylation alters protein activity: blockade of G(o) stimulation by GAP-43. EMBO J. 1992 Jun;11(6):2095–2102. doi: 10.1002/j.1460-2075.1992.tb05268.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Suidan H. S., Stone S. R., Hemmings B. A., Monard D. Thrombin causes neurite retraction in neuronal cells through activation of cell surface receptors. Neuron. 1992 Feb;8(2):363–375. doi: 10.1016/0896-6273(92)90302-t. [DOI] [PubMed] [Google Scholar]
  46. Van Hooff C. O., Holthuis J. C., Oestreicher A. B., Boonstra J., De Graan P. N., Gispen W. H. Nerve growth factor-induced changes in the intracellular localization of the protein kinase C substrate B-50 in pheochromocytoma PC12 cells. J Cell Biol. 1989 Mar;108(3):1115–1125. doi: 10.1083/jcb.108.3.1115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Yankner B. A., Benowitz L. I., Villa-Komaroff L., Neve R. L. Transfection of PC12 cells with the human GAP-43 gene: effects on neurite outgrowth and regeneration. Brain Res Mol Brain Res. 1990 Jan;7(1):39–44. doi: 10.1016/0169-328x(90)90071-k. [DOI] [PubMed] [Google Scholar]
  48. Zuber M. X., Goodman D. W., Karns L. R., Fishman M. C. The neuronal growth-associated protein GAP-43 induces filopodia in non-neuronal cells. Science. 1989 Jun 9;244(4909):1193–1195. doi: 10.1126/science.2658062. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES