Skip to main content
Download PDF
  • Short Communication
  • Published:

Sodium nitroprusside, a nitric oxide donor, fails to bypass the block of neuronal differentiation in PC12 cells imposed by a dominant negative Ras protein

Cellular & Molecular Biology Letters volume 17, pages 323–332 (2012)Cite this article

Abstract

Nitric oxide (NO) is a mediator of a diverse array of inter- and intracellular signal transduction processes. The aim of the present study was to analyze its possible role as a second messenger in the process of neuronal differentiation of PC12 pheochromocytoma cells. Upon NGF treatment wildtype PC12 cells stop dividing and develop neurites. In contrast, a PC12 subclone (designated M-M17-26) expressing a dominant-negative mutant Ras protein keeps proliferating and fails to grow neurites after NGF treatment. Sodium nitroprusside (SNP), an NO donor, was found to induce the p53 protein and to inhibit proliferation of both PC12 and M-M17-26 cells, but failed to induce neuronal differentiation in these cell lines. Key signaling pathways (the ERK and Akt pathways) were also not affected by SNP treatment, and the phosphorylation of CREB transcription factor was only slightly stimulated. It is thus concluded from the results presented in this paper that NO is unable to activate signaling proteins acting downstream or independent of Ras that are required for neuronal differentiation.

Abbreviations

cGMP:

cyclic guanosine monophosphate

CREB:

cAMPresponsive element binding protein

ERK:

extracellular signal-regulated kinase

MAPK:

mitogen-activated protein kinase

NGF:

nerve growth factor

NO:

nitric oxide

NOS:

nitric oxide synthase

PI3K:

phosphatidylinositol 3-kinase

SNP:

sodium nitroprusside

References

  1. Contestabile, A. and Ciani, E. Role of nitric oxide in the regulation of neuronal proliferation, survival and differentiation. Neurochem. Int. 45 (2004) 903–914.

    Article  PubMed  CAS  Google Scholar 

  2. Blaise, G.A., Gauvin, D., Gangal, M. and Authier, S. Nitric oxide, cell signaling and cell death. Toxicology 208 (2005) 177–192.

    Article  CAS  Google Scholar 

  3. Phung, Y.T., Bekker, J.M., Hallmark, O.G. and Black, S.M. Both neuronal NO synthase and nitric oxide are required for PC12 cell differentiation: a cGMP independent pathway. Brain Res. Mol. Brain Res. 64 (1999) 165–178.

    Article  PubMed  CAS  Google Scholar 

  4. Kalisch, B.E., Demeris, C.S., Ishak, M. and Rylett, R.J. Modulation of nerve growth factor-induced activation of MAP kinase in PC12 cells by inhibitors of nitric oxide synthase. J. Neurochem. 87 (2003) 1321–1332.

    Article  PubMed  CAS  Google Scholar 

  5. Martinez-Ruiz, A. and Lamas, S. S-nitrosylation: a potential new paradigm in signal transduction. Cardiovasc. Res. 62 (2004) 43–52.

    Article  PubMed  CAS  Google Scholar 

  6. Yamakura, F. and Ikeda, K. Modification of tryptophan and tryptophan residues in proteins by reactive nitric oxide species. Nitric Oxide 14 (2006) 152–161.

    Article  PubMed  CAS  Google Scholar 

  7. Abello, A., Kerstjens, H.A., Postma, D.S. and Bischoff, R. Protein tyrosine nitration: selectivity, physicochemical and biological consequences, denitration, and proteomics methods for the identification of tyrosine-nitrated proteins. J. Proteome Res. 8 (2009) 3222–3238.

    Article  PubMed  CAS  Google Scholar 

  8. Mikkelsen, R.B. and Wardman, P. Biological chemistry of reactive oxygen and nitrogen and radiation-induced signal transduction mechanisms. Oncogene 22 (2003) 5734–5754.

    Article  PubMed  CAS  Google Scholar 

  9. Greene, L.A. and Tischler, A.S. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc. Natl. Acad. Sci. USA 73 (1976) 2424–2428.

    Article  PubMed  CAS  Google Scholar 

  10. Szeberényi, J. and Erhardt, P. Cellular components of nerve growth factor signaling. Biochim. Biophys. Acta 1222 (1994) 187–202.

    Article  PubMed  Google Scholar 

  11. Greene, L.A. and Angelastro, J.M. You can’t go home again: transcriptionally driven alteration of cell signaling by NGF. Neurochem. Res. 30 (2005) 1347–1352.

    Article  PubMed  CAS  Google Scholar 

  12. Wood, K.W., Sarnecki, C., Roberts, T.M. and Blenis, J. ras mediates nerve growth factor receptor modulation of three signal-transducing protein kinases: MAP kinase, Raf-1 and Rsk. Cell 68 (1992) 1041–1050.

    Article  PubMed  CAS  Google Scholar 

  13. Schonhoff, C.M., Bulseco, D.A., Brancho, D.M., Parada, L.F. and Ross, A.H. The Ras-ERK pathway is required for the induction of neuronal nitric oxide synthase in differentiating PC12 cells. J. Neurochem. 78 (2001) 631–639.

    Article  PubMed  CAS  Google Scholar 

  14. Peunova, N. and Enikolopov, G. Nitric oxide triggers a switch to growth arrest during differentiation of neuronal cells. Nature 375 (1995) 68–73.

    Article  PubMed  CAS  Google Scholar 

  15. Poluha, W., Schonhoff, C.M., Harrington K.S., Lachyankar M.B., Crosbie N.E., Bulseco D.A. and Ross, A.H. A novel, nerve growth factor-activated pathway involving nitric oxide, p53, and p21WAF1 regulates neuronal differentiation of PC12 cells. J. Biol. Chem. 272 (1997) 24002–24007.

    Article  PubMed  CAS  Google Scholar 

  16. Szeberényi, J., Cai, H. and Cooper, G.M. Effect of a dominant inhibitory Ha-Ras mutation on neuronal differentiation of PC12 cells. Mol. Cell. Biol. 10 (1990) 5324–5332.

    PubMed  Google Scholar 

  17. Erhardt, P., Troppmair, J., Rapp, U.R. and Cooper, G.M. Differential regulation of Raf1 and B-RAF and Ras-dependent activation of mitogenactivated protein kinase by cyclic AMP in PC12 cells. Mol. Cell. Biol. 15 (1995) 5524–5530.

    PubMed  CAS  Google Scholar 

  18. Boglári, G. Erhardt, P., Cooper, G.M. and Szeberényi, J. Intact Ras function is required for sustained activation and nuclear translocation of extracellular signal-regulated kinases in nerve growth factor stimulated PC12 cells. Eur. J. Cell. Biol. 75 (1998) 54–58.

    Article  PubMed  Google Scholar 

  19. Kiss, K., Bartek, B., Nusser, N. and Szeberényi, J. Ras-dependence of nerve growth factor-induced inhibition of proliferation of PC12 cells. Acta Biol. Hung. 49 (1998) 97–102.

    CAS  Google Scholar 

  20. Marampon, F., Casimiro, M.C., Fu, M., Powell, M. J., Popov, V.M., Lindsay, J., Zani, B.M., Ciccarelli, C., Watanabe, G., Lee, R.J. and Pestell, R.G. Nerve growth factor regulation of Cyclin D1 in PC12 cells through a p21RAS extracellular signal-regulated kinase pathway requires cooperative interactions between Sp1 and nuclear factor-κB. Mol. Biol. Cell 19 (2008) 2566–2578.

    Article  PubMed  CAS  Google Scholar 

  21. Szeberényi, J., Erhardt, P., Cai, H. and Cooper, G.M. Role of Ras in signal transduction from the nerve growth factor receptor: relationship to protein kinase C, calcium and cyclic AMP. Oncogene 7 (1992) 2105–2113.

    PubMed  Google Scholar 

  22. Boglári, G. and Szeberényi, J. Nuclear translocation of p90Rsk and phosphorylation of CREB is induced by ionomycin in a Ras-independent manner in PC12 cells. Acta Biol. Hung. 53 (2002) 325–334.

    Article  PubMed  Google Scholar 

  23. Du K. and Montminy M. CREB is a regulatory target for the protein kinase Akt/PKB. J. Biol. Chem. 273 (1998) 32377–32379.

    Article  PubMed  CAS  Google Scholar 

  24. De Cesare D., Fimia G.M. and Sassone-Corsi P. Signaling routes to CREM and CREB: plasticity in transcriptional activation. Trends Biochem. Sci. 24 (1999) 281–285.

    Article  PubMed  Google Scholar 

  25. Lander, H.M., Jacovina, A.T., Davis, R.J. and Tauras, J.M. Differential activation of mitogen-activated protein kinases by nitric oxide-related species. J. Biol. Chem. 271 (1996) 19705–19709.

    Article  PubMed  CAS  Google Scholar 

  26. Ciani, E., Guidi, S., Bartesaghi, R. and Contestabile, A. Nitric oxide regulates cGMP-dependent cAMP-responsive element binding protein phosphorylation and Bcl-2 expression in cerebellar neurons: implication for a survival role of nitric oxide. J. Neurochem. 82 (2002) 1282–1289.

    Article  PubMed  CAS  Google Scholar 

  27. Rudkin, B.B., Lazarovici, P., Levi, B.Z., Abe, Y., Fujita, K. and Guroff, G. Cell cycle-specific action of nerve growth factor in PC12 cells: differentiation without proliferation. EMBO J. 8 (1989) 3319–3325.

    PubMed  CAS  Google Scholar 

  28. Brynczka, C. and Merrick, B.A. Nerve growth factor potentiates p53 DNA binding but inhibits nitric oxide-induced apoptosis in neuronal PC12 cells. Neurochem. Res. 32 (2007) 1573–1585.

    Article  PubMed  CAS  Google Scholar 

  29. Jänicke, R.U., Graupner, V., Budach, W., Essmann, F. The do’s and don’ts of p53 isoforms. Biol. Chem. 390 (2009) 951–963.

    Article  PubMed  Google Scholar 

  30. Hindley, S., Juurlink, B.H., Gysbers, J.W., Middlemiss, P.J., Herman, M.A. and Rathbone M.P. Nitric oxide donors enhance neurotrophininduced neurite outgrowth through a cGMP-dependent mechanism. J. Neurosci. Res. 47 (1997) 427–439.

    Article  PubMed  CAS  Google Scholar 

  31. Li, C.Q. and Wogan, G.N. Nitric oxide as a modulator of apoptosis. Cancer Lett. 226 (2005) 1–15.

    Article  PubMed  CAS  Google Scholar 

  32. Traverse, S., Gomez, N., Paterson, H., Marshall, C.J. and Cohen, P. Sustained activation of the mitogen-activated protein (MAP) kinase cascade may be required for differentiation of PC12 cells. Biochem. J. 288 (1992) 351–355.

    PubMed  CAS  Google Scholar 

  33. Lander, H.M., Hajjar, D.P., Hempstead, B.L., Mirza, U.A., Chait, B.T., Campbell, S. and Quilliam, L.A. A molecular redox switch on p21(ras). Structural basis for the nitric oxide-p21(ras) interaction. J. Biol. Chem. 272 (1997) 4323–4326.

    Article  PubMed  CAS  Google Scholar 

  34. Heo, J. and Campbell, S.L. Mechanism of p21Ras S-nitrosylation and kinetics of nitric oxide-mediated guanine nucleotide exchange. Biochemistry 43 (2004) 2314–2322.

    Article  PubMed  CAS  Google Scholar 

  35. Jeong, H.S., Kim, S.W., Baek, K.J., Lee, H.S., Kwon, N.S., Kim, Y.M. and Yun, H.Y. Involvement of Ras in survival responsiveness to nitric oxide toxicity in pheochromocytoma cells. J. Neurooncol. 60 (2002) 97–107.

    Article  PubMed  Google Scholar 

  36. Yao, R. and Cooper G.M. Requirement of phosphatidylinositol 3-kinase in the prevention of apoptosis by nerve growth factor. Science 267 (1995) 2003–2006.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Medical Biology, Medical School, University of Pécs, 7624, Pécs, Szigeti u. 12, Hungary

    Judit Bátor, Judit Varga, Gergely Berta & József Szeberényi

  2. Ilia State University, Tbilisi, Georgia

    Tamar Barbakadze & David Mikeladze

  3. Collegium Basilea, Institute of Advanced Study, Basel, Switzerland

    Jeremy Ramsden

Authors
  1. Judit Bátor
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Judit Varga
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gergely Berta
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tamar Barbakadze
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. David Mikeladze
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jeremy Ramsden
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. József Szeberényi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to József Szeberényi.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bátor, J., Varga, J., Berta, G. et al. Sodium nitroprusside, a nitric oxide donor, fails to bypass the block of neuronal differentiation in PC12 cells imposed by a dominant negative Ras protein. Cell Mol Biol Lett 17, 323–332 (2012). https://doi.org/10.2478/s11658-012-0013-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11658-012-0013-8

Key words

Cellular & Molecular Biology Letters

ISSN: 1689-1392

Contact us