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. 1988 Nov;85(21):8296–8300. doi: 10.1073/pnas.85.21.8296

The 43-kDa neuronal growth-associated protein (GAP-43) is present in plasma membranes of rat astrocytes.

L Vitković 1, H W Steisslinger 1, V J Aloyo 1, M Mersel 1
PMCID: PMC282416  PMID: 3054883

Abstract

One of the neuronal growth-associated proteins, GAP-43 (molecular mass, approximately 43 kDa; pI 4.3), is abundant in growth-cone membranes and corresponds to a major protein kinase C substrate, the 46-kDa phosphoprotein (pp46), of a growth-cone-enriched subcellular fraction. This protein has the following additional designations (depending on context): B-50 (phospholipid metabolism), F1 (synaptic plasticity), and p57 (calmodulin binding). We show that a protein with the same molecular mass and isoelectric point as GAP-43, which interacts with anti-GAP-43 antibodies on immunoblots, is present in the plasma membranes of cultured neonatal rat cortical astrocytes. Double-immunofluorescence labeling of cells with a serum against glial fibrillary acidic protein and anti-GAP-43 antibody was observed. Furthermore, astrocytic protein was phosphorylated in vitro by protein kinase C and comigrated in two-dimensional PAGE with GAP-43. The data indicate that GAP-43, heretofore believed to be neuron-specific, is present in at least one class of glial cells.

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Selected References

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

  1. Aloyo V. J., Zwiers H., Gispen W. H. B-50 protein kinase and kinase C in rat brain. Prog Brain Res. 1982;56:303–315. doi: 10.1016/S0079-6123(08)63781-4. [DOI] [PubMed] [Google Scholar]
  2. Aronson N. N., Jr, Touster O. Isolation of rat liver plasma membrane fragments in isotonic sucrose. Methods Enzymol. 1974;31:90–102. doi: 10.1016/0076-6879(74)31009-9. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Benowitz L. I., Perrone-Bizzozero N. I., Finklestein S. P. Molecular properties of the growth-associated protein GAP-43 (B-50). J Neurochem. 1987 May;48(5):1640–1647. doi: 10.1111/j.1471-4159.1987.tb05713.x. [DOI] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Crane F. L., Löw H. NADH oxidation in liver and fat cell plasma membranes. FEBS Lett. 1976 Oct 1;68(2):153–156. doi: 10.1016/0014-5793(76)80425-5. [DOI] [PubMed] [Google Scholar]
  7. Ellis L., Wallis I., Abreu E., Pfenninger K. H. Nerve growth cones isolated from fetal rat brain. IV. Preparation of a membrane subfraction and identification of a membrane glycoprotein expressed on sprouting neurons. J Cell Biol. 1985 Nov;101(5 Pt 1):1977–1989. doi: 10.1083/jcb.101.5.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fischel S. V., Medzihradsky F. Assessment of membrane permeability in primary cultures of neurons and glia in response to osmotic perturbation. J Neurosci Res. 1985;13(3):369–380. doi: 10.1002/jnr.490130304. [DOI] [PubMed] [Google Scholar]
  9. Fowler C. J., Tipton K. F. Concentration dependence of the oxidation of tyramine by the two forms of rat liver mitochondrial monoamine oxidase. Biochem Pharmacol. 1981 Dec 15;30(24):3329–3332. doi: 10.1016/0006-2952(81)90607-9. [DOI] [PubMed] [Google Scholar]
  10. Hubbard A. L., Cohn Z. A. Externally disposed plasma membrane proteins. I. Enzymatic iodination of mouse L cells. J Cell Biol. 1975 Feb;64(2):438–460. doi: 10.1083/jcb.64.2.438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jacobson R. D., Virág I., Skene J. H. A protein associated with axon growth, GAP-43, is widely distributed and developmentally regulated in rat CNS. J Neurosci. 1986 Jun;6(6):1843–1855. doi: 10.1523/JNEUROSCI.06-06-01843.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Karns L. R., Ng S. C., Freeman J. A., Fishman M. C. Cloning of complementary DNA for GAP-43, a neuronal growth-related protein. Science. 1987 May 1;236(4801):597–600. doi: 10.1126/science.2437653. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Mersel M., Malviya A. N., Hindelang C., Mandel P. Plasma membrane isolated from astrocytes in primary cultures. Its acceptor oxidoreductase properties. Biochim Biophys Acta. 1984 Nov 21;778(1):144–154. doi: 10.1016/0005-2736(84)90458-9. [DOI] [PubMed] [Google Scholar]
  15. Mersel M., Tholey G., Delaunoy J. P., Rebel G., Flory E., Mandel P. Isolement et caractérisation d'une nouvelle lignée d'astrocytes spontanément transformés du cerveau de rat. C R Acad Sci III. 1985;301(19):811–815. [PubMed] [Google Scholar]
  16. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  17. Neary J. T., Norenberg L. O., Norenberg M. D. Protein kinase C in primary astrocyte cultures: cytoplasmic localization and translocation by a phorbol ester. J Neurochem. 1988 Apr;50(4):1179–1184. doi: 10.1111/j.1471-4159.1988.tb10590.x. [DOI] [PubMed] [Google Scholar]
  18. Neve R. L., Perrone-Bizzozero N. I., Finklestein S., Zwiers H., Bird E., Kurnit D. M., Benowitz L. I. The neuronal growth-associated protein GAP-43 (B-50, F1): neuronal specificity, developmental regulation and regional distribution of the human and rat mRNAs. Brain Res. 1987 Jul;388(2):177–183. doi: 10.1016/s0006-8993(87)80012-4. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  21. Perrone-Bizzozero N. I., Finklestein S. P., Benowitz L. I. Synthesis of a growth-associated protein by embryonic rat cerebrocortical neurons in vitro. J Neurosci. 1986 Dec;6(12):3721–3730. doi: 10.1523/JNEUROSCI.06-12-03721.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pfenninger K. H., Ellis L., Johnson M. P., Friedman L. B., Somlo S. Nerve growth cones isolated from fetal rat brain: subcellular fractionation and characterization. Cell. 1983 Dec;35(2 Pt 1):573–584. doi: 10.1016/0092-8674(83)90191-5. [DOI] [PubMed] [Google Scholar]
  23. Sensenbrenner M., Devilliers G., Bock E., Porte A. Biochemical and ultrastructural studies of cultured rat astroglial cells: effect of brain extract and dibutyryl cyclic AMP on glial fibrillary acidic protein and glial filaments. Differentiation. 1980;17(1):51–61. doi: 10.1111/j.1432-0436.1980.tb01081.x. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Skene J. H., Willard M. Changes in axonally transported proteins during axon regeneration in toad retinal ganglion cells. J Cell Biol. 1981 Apr;89(1):86–95. doi: 10.1083/jcb.89.1.86. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Steisslinger H. W., Aloyo V. J., Vitković L. Characterization of two plasma membrane proteins abundant in rat brain. Brain Res. 1987 Jul 14;415(2):375–379. doi: 10.1016/0006-8993(87)90224-1. [DOI] [PubMed] [Google Scholar]
  27. Temple S., Raff M. C. Differentiation of a bipotential glial progenitor cell in a single cell microculture. Nature. 1985 Jan 17;313(5999):223–225. doi: 10.1038/313223a0. [DOI] [PubMed] [Google Scholar]
  28. Zwiers H., Verhaagen J., van Dongen C. J., de Graan P. N., Gispen W. H. Resolution of rat brain synaptic phosphoprotein B-50 into multiple forms by two-dimensional electrophoresis: evidence for multisite phosphorylation. J Neurochem. 1985 Apr;44(4):1083–1090. doi: 10.1111/j.1471-4159.1985.tb08728.x. [DOI] [PubMed] [Google Scholar]

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