Nerve Growth Factor Activates Brain-derived Neurotrophic Factor Promoter IV via Extracellular Signal-regulated Protein Kinase 1/2 in PC12 Cells

  • Park, So Yun (School of Biological Sciences, Seoul National University) ;
  • Lee, Ji Yun (Department of Anatomy, School of Medicine, Kyungpook National University) ;
  • Choi, Jun Young (Department of Anatomy, School of Medicine, Kyungpook National University) ;
  • Park, Mae Ja (Department of Anatomy, School of Medicine, Kyungpook National University) ;
  • Kim, Dong Sun (Department of Anatomy, School of Medicine, Kyungpook National University)
  • Received : 2005.12.07
  • Accepted : 2006.01.18
  • Published : 2006.04.30


Brain-derived neurotrophic factor (BDNF) is a neuromodulator of nociceptive responses in the dorsal root ganglia (DRG) and spinal cord. BDNF synthesis increases in response to nerve growth factor (NGF) in trkA-expressing small and medium-sized DRG neurons after inflammation. Previously we demonstrated differential activation of multiple BDNF promoters in the DRG following peripheral nerve injury and inflammation. Using reporter constructs containing individual promoter regions, we investigated the effect of NGF on the multiple BDNF promoters, and the signaling pathway by which NGF activates these promoters in PC12 cells. Although all the promoters were activated 2.4-7.1-fold by NGF treatment, promoter IV gave the greatest induction. The p38 mitogen-activated protein kinase (MAPK) inhibitor, SB203580, phosphatidylinositol 3-kinase (PI-3K) inhibitor, LY294003, protein kinase A (PKA) inhibitor, H89, and protein kinase C (PKC) inhibitor, chelerythrine, had no effect on activation of promoter IV by NGF. However, activation was completely abolished by the MAPK kinase (MEK) inhibitors, U0126 and PD98059. In addition, these inhibitors blocked NGF-induced phosphorylation of extracellular signal-regulated protein kinase (ERK) 1/2. Taken together, these results suggest that the ERK1/2 pathway activates BDNF promoter IV in response to NGF independently of NGF-activated signaling pathways involving PKA and PKC.


BDNF;DRG;ERK1/2;Inflammatory Pain;Multiple Promoters;NGF;PC12 Cells


Supported by : Korea Research Foundation


  1. Aley, K. O. and Levine, J. D. (1999) Role of protein kinase A in the maintenance of inflammatory pain. J. Neurosci. 19, 2181-2186
  2. Aley, K. O., Martin, A., McMahon, T., Mok, J., Levine, J. D., et al. (2001) Nociceptor sensitization by extracellular signalregulated kinases. J. Neurosci. 21, 6933-6939
  3. Doya, H., Ohtori, S., Fujitani, M., Saito, T., Hata, K., et al. (2005) c-Jun N-terminal kinase activation in dorsal root ganglion contributes to pain hypersensitivity. Biochem. Biophys. Res. Commun. 335, 132-138
  4. Landry, J. R., Mager, D. L., and Wilhelm, B. T. (2003) Complex controls: the role of alternative promoters in mammalian genomes. Trends Genet. 19, 640-648
  5. Liu, Y. Z., Chirivia, J. C., and Latchman, D. S. (1998) Nerve growth factor up-regulates the transcriptional activity of CBP through activation of the p42/p44(MAPK) cascade. J. Biol. Chem. 273, 32400-32407
  6. Michael, G. J., Averill, S., Shortland, P. J., Yan, Q., and Priestley, J. V. (1999) Axotomy results in major changes in BDNF expression by dorsal root ganglion cells: BDNF expression in large trkB and trkC cells, in pericellular baskets, and in projection to deep dorsal horn and dorsal column nuclei. Eur. J. Neurosci. 11, 3539-3551
  7. Nakayama, M., Gahara, Y., Kitamura, T., and Ohara, D. (1994) Distinctive four promoters collectively direct expression of brain-derived neurotrophic factor gene. Brain Res. Mol. Brain Res. 21, 206-218
  8. Obata, K., Yamanaka, H., Dai, Y., Mizushima, T., Fukuoka, T., et al. (2004a) Activation of extracellular signal-regulated protein kinase in the dorsal root ganglion following inflammation near the nerve cell body. Neuroscience 126, 1011-1021
  9. Shieh, P. B., Hu, S. C., Bobb, K., Timmusk, T., and Ghosh, A. (1998) Identification of a signaling pathway involved in calcium regulation of BDNF expression. Neuron 20, 727-740
  10. Thoenen, H. (1995) Neurotrophins and neuronal plasticity. Science 270, 593-598
  11. Kim, D. S., Lee, S. J., and Cho, H. J. (2001) Differential usage of multiple brain-derived neurotrophic factor promoter in rat dorsal root ganglia following peripheral nerve injuries and inflammation. Brain Res. Mol. Brain Res. 92, 167-171
  12. Michael, G. J., Averill, S., Nitkunan, A., Rattray, M., Bennett, D. L., et al. (1997) Nerve growth factor treatment increases brain-derived neurotrophic factor selectively in TrkA-expressing dorsal root ganglion cells and their centrl terminations within the spinal cord. J. Neurosci. 17, 8476-8490
  13. Watson, A. and Latchman, D. S. (1995) The cyclic AMP response element in the calcitonin/calcitonin gene-related peptide gene promoter is necessary but not sufficient for its activation by nerve growth factor. J. Biol. Chem. 270, 9655-9660
  14. Obata, K. and Noguchi, K. (2004) MAPK activation in nociceptive neurons and pain hypersensitivity. Life Sci. 74, 2643-2653
  15. Delcroix, J. D., Valletta, J. S., Wu, C., Hunt, S. J., Kowal, A. S., et al. (2003) NGF signaling in sensory neurons: evidence that early endosomes carry NGF retrograde signals. Neuron 39, 69-84
  16. Averill, S., Delcroix, J. D., Michael, G. J., Tomlinson, D. R., Fernyhough, P., et al. (2001) Nerve growth factor modulates the activation status and fast axonal transport of erk 1/2 in adult nociceptive neurons. Mol. Cell Neurosci. 18, 183-196
  17. Metsis, M., Timmusk, T., Arenas, E., and Persson, H. (1993) Differential usage of multiple brain-derived neurotrophic factor promoters in the rat brain following neuronal activation. Proc. Natl. Acad. Sci. USA 90, 8802-8806
  18. Brons, R., Klesse, L. J., Shah, K., Parada, L. F., Winter, J. (2003) Activation of Ras is necessary and sufficient for upregulation of vanilloid receptor type 1 in sensory neurons by neurotrophic factors. Mol. Cell Neurosci. 22, 118-132
  19. Ji, R. R. (2004) Mitogen-activated protein kinases as a potential targets for pain killers. Curr. Opin. Investig. Drugs 5, 71- 75
  20. Ferreira, J., Triches, K. M., Medeiros, R., and Calixto, J. B. (2005) Mechanisms involved in the nociception produced by peripheral protein kinase C activation in mice. Pain 117, 171-181
  21. Obata, K., Yamanaka, H., Dai, Y., Tachibana, T., Fukuoka, T., et al. (2004b) Differential activation of extracellular signalregulated protein kinase in primary afferent neurons regulates brain-derived neurotrophic factor expression after peripheral inflammation and nerve injury. J. Neurosci. 23, 4117-4126
  22. Woolf, C. J. and Costigan, M. (1999) Transcriptional and posttranslational plasticity and the generation of inflammatory pain. Proc. Natl. Acad. Sci. USA 96, 7723-7730
  23. Dai, Y., Iwata, K., Fukuoka, T., Kondo, E., Tokunaga, A., et al. (2002) Phosphorylation of extracellular signal-regulated kinase in primary afferent neurons by noxious stimuli and its involvemnet in peripheral sensitization. J. Neurosci. 22, 7737-7745
  24. Kokaia, Z., Metsis, M., Kokaia, M., Bengzon, J., Elmer, E., et al. (1994) Brain insults in rats induce increased expression of the BDNF gene through differential use of multiple promoters. Eur. J. Neurosci. 6, 587-596
  25. Cho, H. J., Kim, J. K., Park, H. C., Kim, J. K., Kim, D. S., et al. (1998) Changes in brain-derived neurotrophic factor immunoreactivity in rat dorsal root ganglia, spinal cord, and gracile nuclei following cut or crush injuries. Exp. Neurol. 154, 224-230
  26. Ji, R. R., Baba, H., Brenner, G. J., and Woolf, C. J. (1999) Nociceptive- specific activation of ERK in spinal neurons contributes to pain hypersensitivity. Nat. Neurosci. 2, 1114-1119
  27. Sweatt, J. D. (2001) The neuronal MAP kinase cascade: a biochemical signal integration system subserving synaptic plasticity and memory. J. Neurochem. 76, 1-10
  28. Ji, R. R., Befort, K., Brenner, G. J., and Woolf, C. J. (2002a) ERK MAP kinase activation in superficial spinal cord neurons induces prodynorphin and NK-1 upregulation and contributes to persistent inflammatory pain hypersensitivity. J. Neurosci. 22, 478-485
  29. Kim, S. Y., Bae, J. C., Kim, J. Y., Lee, H. L., Lee, M., et al. (2002) Activation of p38 Map kinase in the rat dorsal root ganglia and spinal cord following peripheral inflammation and nerve injury. Neuroreport 13, 2483-2486
  30. Liu, Y. Z., Thomas, N. S. B., and Latchman, D. S. (1999) CBP associates with the p42/p44 MAPK enzymes and is phosphorylated following NGF treatment. Neuroreport 10, 1239-1243
  31. Thomson, S. W., Bennett, D. L. H., Kerr, B. J., Bradbury, E. J., and McMahon, S. B. (1999) Brain-derived neurotrophic factor is an endogenous modulator of nociceptive responses in the spinal cord. Proc. Natl. Acad. Sci. USA 96, 7714-7718
  32. Millan, M. J. (1999) The introduction of pain: an integrative review. Prog. Neurobiol. 57, 1-164
  33. Schaeffer, H. J. and Weber, M. J. (1999) Mitogen-activated protein kinases: specific messages from ubiquitous messengers. Mol. Cell. Biol. 19, 2435-2444
  34. Oliff, H. S., Berchtold, N. C., Isackson, P., and Cotman, C. W. (1998) Exercise-induced regulation of brain-derived neurotrophic factor (BDNF) transcripts in the rat hippocampus. Brain Res. Mol. Brain Res. 61, 147-153
  35. Slagsvold, H. H., Ostvold, A. C., Fallgren, A. B., and Paulsen, R. E. (2002) Nuclear recptor and apoptosis initiator NGFI-B is a substrate for kinase ERK2. Biochem. Biophys. Res. Commun. 291, 1146-1150
  36. Apfel, S. C., Wright, D. E., Wiideman, A. M., Dormia, C., Snider, W. D., et al. (1996) Nerve growth factor regulates the expression of brain-derived neurotrophic factor mRNA in the peripheral nerve system. Mol. Cell Neurosci. 7, 134-142
  37. Dina, O. A., McCarter, G. C., de Coupade, C., and Levine, J. D. (2003) Role of sensory neuron cytoskleton in second messenger signaling for inflammatory pain. Neuron 39, 613-624
  38. Khasar, S. G., Lin, Y. H., Martin, A., Dadgar, J., McMahon, T., et al. (1999) A novel nociceptor signaling pathway revealed in protein kinase C epsilon mutant mice. Neuron 24, 253-260
  39. Pezet, S., Malcangio, M., and McMahon, S. B. (2002) BDNF: a neuromodulator in nociceptive pathways. Brain Res. Rev. 40, 240-249
  40. Timmusk, T., Palm, K., Medis, M., Reintam, T., Paalme, V., et al. (1993) Multiple promoters direct tissue-specific expression of the rat BDNF gene. Neuron 10, 475-489
  41. Tabuchi, A., Nakaoka, R., Amano, K., Yukimine, M., Andoh, T., et al. (2000) Differential activation of brain-derived neurotrophic factor gene promoters I and III by $Ca^{2+}^ signals evoked via L-type voltage-dependent and N-methyl-Daspartate receptor $Ca^{2+}^ channels. J. Biol. Chem. 275, 17269- 17275
  42. Takeuchi, Y., Miyamoto, E., and Fukunaga, K. (2003) Analysis on the promoter region of exon IV brain-derived neurotrophic factor in NG108-15 cells. J. Neurochem. 83, 67-79
  43. Ji, R. R., Samad, T. A., Jin, S. X., Schmoll, R., and Woolf, C. J. (2002b) p38 MAPK activation in primary sensory neurons after inflammation increase TRPV1 levels and maintains heat hyperalgesia. Neuron 36, 57-68
  44. Nanda, S. and Mack, K. J. (1998) Multiple promoters direct stimulus and temporal specific expression of brain-derived neurotrophic actor in the somatosensory cortex. Brain Res. Mol. Brain Res. 62, 216-219
  45. Tabuchi, A., Sakaya, H., Kisukeda, T., Fukuchi, M., and Tsuda, M. (2002) Involvement of an upstream stimulatory factor as well as cAMP-responsive element-binding protein in the activation of brain-derived neurotrophic factor gene promoter I. J. Biol. Chem. 277, 35920-31
  46. Xing, J., Ginty, D. D., and Greenberg, M. E. (1996) Coupling of the RAS-MAPK pathway to gene activation by RSK2, a growth factor-regulated CREB kinase. Science 273, 959-963
  47. Zha, X. M., Bishop, J. F., Hansen, M. R., Victoria, L., Abbas, P. J., et al. (2001) BDNF synthesis in spiral ganglion neurons in constitutive and CREB-dependent. Hear. Res. 156, 53-68
  48. Freeland, K., Liu, Y. Z., and Latchman, D. S. (2000) Distinct signalling pathways mediate the cAMP response element (CRE)-dependent activation of the calcitonin gene-related peptide gene promoter by cAMP and nerve growth factor. Biochem. J. 345, 233-238
  49. Bishop, J. F., Mueller, G. P., and Mouradian, M. M. (1994) Alternative 5′ exons in the rat brain-derived neurotrophic factor gene: differential patterns of expression across brain regions. Brain Res. Mol. Brain Res. 26, 225-232