Identification of two nerve growth factor-induced polypeptides in PC12 cells

  • Mark A. Sussman
  • Elena Battenberg
  • Floyd E. Bloom
  • Velia M. Fowler


We have identified two nerve growth factor (NGF)-induced polypeptides (Mr 80,000 and 90,000) in PC12 cells that are heat stable and not sulfated. Indirect immunofluorescence localization of these polypeptides in NGF-treated PC12 cells reveals a punctate pattern concentrated in neurites. Immunoperoxidase staining of rat tissue shows that these polypeptides are found throughout the brain in selected subsets of neurons and are absent in the pituitary or adrenal medulla. Several of these characteristics are similar to a recently described NGF-induced secretory protein (VGF8a), which has sequence similarities to secretogranins. The property of NGF inducibility and the distribution of these polypeptides within rat tissues are both novel features for secretogranin proteins.


PC12 Cell Nerve Growth Factor Nerve Growth Factor Treatment Antipeptide Antibody Nitrocellulose Strip 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. Anderson, D.J., Axel, R. (1985). Molecular probes for the development and plasticity of neural crest derivatives. Cell 42:649–662PubMedCrossRefGoogle Scholar
  2. Bloom, F.E., Battenberg, E.L.F. (1983). Immunocytochemistry of endorphins and enkephalins. Methods Enzymol. 103:670–687PubMedCrossRefGoogle Scholar
  3. Bottenstein, J.E., Sato, G.H. (1979). Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc. Natl. Acad. Sci. U.S.A. 76:514–517PubMedCrossRefGoogle Scholar
  4. Brugg, B., Matus, A. (1988). PC12 cells express juvenile microtubule-associated proteins during nerve growth factor-induced neurite outgrowth. J. Cell Biol. 107:643–650PubMedCrossRefGoogle Scholar
  5. Burke, B., Griffiths, G., Reggio, H., Louvard, D., Warren, G. (1982). A monoclonal antibody against a 135K Golgi membrane protein. EMBO J. 1:1621–1628PubMedGoogle Scholar
  6. Dichter, M.A., Tischler, A.S., Greene, L.A. (1977). Nerve growth factor-induced change in electrical excitability and acetylcholine sensitivity of a rat pheochromocytoma cell line. Nature (London) 268:501–504CrossRefGoogle Scholar
  7. Dickson, G., Prentice, H., Julien, J.P., Ferrari, G., Leon, A., Walsh, F.S. (1986). Nerve growth factor activates Thy-1 and neurofilament gene transcription in rat PC12 cells. EMBO J. 5:3349–3353Google Scholar
  8. Devereux, J., Haeberli, P., Smithies, O. (1984). A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 12:387–395PubMedCrossRefGoogle Scholar
  9. Doherty, P., Mann, D.A., Walsh, F.S. (1988). Comparison of the effects of NGF, activators of protein kinase C and a calcium ionophore on the expression of Thy-1 and N-CAM in PC12 cell cultures. J. Cell Biol. 107:333–340PubMedCrossRefGoogle Scholar
  10. Fischer-Colbrie, R., Schober, M. (1987). Isolation and characterization of chromogranins A, B, and C from bovine chromaffin granules and a rat pheochromocytoma. J. Neurochem. 48:262–270PubMedCrossRefGoogle Scholar
  11. Forss-Petter, S., Danielson, P., Battenberg, E., Bloom, F., Sutcliffe, J.G. (1989). Nucleotide sequence and cellular distribution of rat chromogranin B (secretogranin I) mRNA in the neuroendocrine system. J. Mol. Neurosci. 1:63–75PubMedGoogle Scholar
  12. Fowler, V.M. (1987). Identification and purification of a novel Mr 43,000 tropomyosin-binding protein from human erythrocyte membranes. J. Biol. Chem. 262:12792–12800PubMedGoogle Scholar
  13. Fowler, V.M. (1990). Tropomodulin: A cytoskeletal protein that binds to the end of erythrocyte tropomyosin and inhibits tropomyosin binding to F-actin. J. Cell Biol. 111:471–482PubMedCrossRefGoogle Scholar
  14. Fowler, V.M., Bennett, V. (1984). Erythrocyte membrane tropomyosin. Purification and properties. J. Biol. Chem. 259:5978–5989PubMedGoogle Scholar
  15. Garrels, J.I., Schubert, D. (1979). Modulation of protein synthesis by nerve growth factor. J. Biol. Chem. 254:7978–7985PubMedGoogle Scholar
  16. Greenberg, M.E., Greene, L.A., Ziff, E.B. (1985). Nerve growth factor and epidermal growth factor induce rapid transient changes in proto-oncogene transcription in PC12 cells. J. Biol. Chem. 260:14101–14110PubMedGoogle Scholar
  17. Greene, L.A., Tischler, A.S. (1976). Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc. Natl. Acad. Sci. U.S.A. 73:2424–2428PubMedCrossRefGoogle Scholar
  18. Greene, L.A., Liem, R.K.H., Shelanski, M.L. (1983). Regulation of a high molecular weight microtubule-associated protein in PC12 cells by nerve growth factor. J. Cell Biol. 96:76–93PubMedCrossRefGoogle Scholar
  19. Ignatius, M.J., Chandler, C.R., Shooter, E.M. (1985). Nerve growth factor-treated, neurite-bearing PC12 cells continue to synthesize DNA. J. Neurosci. 5:343–351PubMedGoogle Scholar
  20. Kreis, T.E. (1986). Micro injected antibodies against the cytoplasmic domain of vesicular stomatitis virus glycoprotein block its transport to the cell surface. EMBO J. 5:931–941PubMedGoogle Scholar
  21. Kruijer, W., Schubert, D., Verma, I.M. (1985). Induction of the proto-oncogenefos by nerve growth factor. Proc. Natl. Acad. Sci. U.S.A. 82:7330–7334PubMedCrossRefGoogle Scholar
  22. Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  23. Lee, R.W.H., Huttner, W.B. (1983). Tyrosine-O-sulfated proteins of PC12 pheochromocytoma cells and their sulfation by tyrosylprotein sulfotransferase. J. Biol. Chem. 258:11326–11334PubMedGoogle Scholar
  24. Leonard, D.G.B., Ziff, E.B., Greene, L.A. (1987). Identification and characterization of mRNAs regulated by nerve growth factor in PC12 cells. Mol. Cell Biol. 7:3156–3167PubMedGoogle Scholar
  25. Letourneau, P.C. (1981). Immunocytochemical evidence for colocalization in neurite growth cones of actin and myosin and their relationship to cell-substratum adhesions. Dev. Biol. 85:113–122PubMedCrossRefGoogle Scholar
  26. Levi, A., Eldridge, J.D., Paterson, B.M. (1985). Molecular cloning of a gene sequence regulated by nerve growth factor. Science 229:393–395PubMedCrossRefGoogle Scholar
  27. Levi-Montalcini, R., Calissano, P. (1986). Nerve growth factor as a paradigm for other polypeptide growth factors. Trends Neurosci. 9:473–477CrossRefGoogle Scholar
  28. Lindenbaum, M.H., Carbonetto, S., Mushynski, W.E. (1987). Nerve growth factor enhances the synthesis, phosphorylation, and metabolic stability of neurofilament proteins in PC12 cells. J. Biol. Chem. 262:605–610PubMedGoogle Scholar
  29. Margolis, R.K., Salton, S.R.J., Margolis, R.U. (1983). Complex carbohydrates of cultured PC12 pheochromocytoma cells. J. Biol. Chem. 258:4110–4117PubMedGoogle Scholar
  30. Masiakowski, P., Shooter, E.M. (1988). Nerve growth factor induces the genes for two proteins related to a family of calcium-binding proteins in PC12 cells. Proc. Natl. Acad. Sci. U.S.A. 85:1277–1281PubMedCrossRefGoogle Scholar
  31. McGuire, J.C., Greene, L.A., Furano, A.V. (1978). NGF stimulates incorporation of fucose or glucosamine into an external glycoprotein in cultured rat pheochromocytoma cells. Cell 15:357–365PubMedCrossRefGoogle Scholar
  32. Milbrandt, J. (1987). A nerve growth factor-induced gene encodes a possible transcriptional regulatory factor. Science 238:797–799PubMedCrossRefGoogle Scholar
  33. Paves, H., Neuman, T., Metsis, M., Saarma, M. (1988). Nerve growth factor induces rapid redistribution of Factin in PC12 cells. FEBS Lett. 235:141–143PubMedCrossRefGoogle Scholar
  34. Possenti, R., Eldridge, J.D., Paterson, B.M., Grasso, A., Levi, A. (1989). A protein induced by NGF in PC12 cells is stored in secretory vesicles and released through the regulated pathway. EMBO J. 8:2217–2223PubMedGoogle Scholar
  35. Richter-Landesberg, C., Lee, V.M., Salton, S.R.J., Shelanski, M.L., Greene, L.A. (1984). Release of the nerve growth factor-inducible large external and other glycoproteins from cultured PC12 rat pheochromocytoma cells and sympathetic neurons. J. Neurochem. 43:841–848CrossRefGoogle Scholar
  36. Rimm, D.L., Pollard, T.D. (1989). Purification and characterization of anAcanthomoeba nuclear actin-binding protein. J. Cell Biol. 109:585–591PubMedCrossRefGoogle Scholar
  37. Rosa, P., Hille, A., Lee, R.W.H., Zanini, A., DeCamilli, P., Huttner, W.B. (1985). Secretogranins I and II: Two tyrosine-sulfated secretory proteins common to a variety of cells secreting peptides by the regulated pathway. J. Cell Biol. 101:1999–2011PubMedCrossRefGoogle Scholar
  38. Salton, S.R.J., Margolis, R.V., Margolis, R.K. (1983). Release of chromaffin granule glycoproteins and proteoglycans from potassium-stimulated PC12 pheochromocytoma cells. J. Neurochem. 41:1165–1170PubMedCrossRefGoogle Scholar
  39. Schubert, D., LaCorbiere, M., Whitlock, C., Stallcup, W. (1978). Alterations in the surface properties of cells responsive to nerve growth factor. Nature 273:718–723PubMedCrossRefGoogle Scholar
  40. Tischler, A.S., Greene, L.A. (1978). Morphologic and cytochemical properties of a clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Lab. Invest. 39:77–89PubMedGoogle Scholar
  41. Towbin, H., Staehelin, T., Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Procedure and some applications. Proc. Natl. Acad. Sci. U.S.A. 76:4350–4354PubMedCrossRefGoogle Scholar
  42. VanHooff, C.O.M., Holthuis, J.C.M., Oestreicher, A.B., Boonstra, J., DeGraan, P.N.E., Gispen, W.H. (1989). Nerve growth factor-induced changes in the intracellular localization of the protein kinase C substrate B-50 in pheochromocytoma PC12 cells. J. Cell Biol. 108:1115–1125CrossRefGoogle Scholar

Copyright information

© Birkhäuser 1990

Authors and Affiliations

  • Mark A. Sussman
    • 1
  • Elena Battenberg
    • 2
  • Floyd E. Bloom
    • 2
  • Velia M. Fowler
    • 1
  1. 1.Department of Molecular BiologyScripps Clinic and Research FoundationLa JollaUSA
  2. 2.Department of NeuropharmacologyScripps Clinic and Research FoundationLa JollaUSA

Personalised recommendations