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. 1998 Aug 1;333(Pt 3):505–509. doi: 10.1042/bj3330505

Stimulation of neuropeptide Y gene expression by brain-derived neurotrophic factor requires both the phospholipase Cgamma and Shc binding sites on its receptor, TrkB.

A G Williams 1, A C Hargreaves 1, F J Gunn-Moore 1, J M Tavaré 1
PMCID: PMC1219610  PMID: 9677306

Abstract

In PC12 cells, it has been previously reported that nerve growth factor stimulates neuropeptide Y (NPY) gene expression. In the current study we examined the signalling pathways involved in this effect by transiently expressing in PC12 cells the receptor (TrkB) for the related neurotrophin, brain-derived neurotrophic factor (BDNF). BDNF caused a 3-fold induction of luciferase expression from a transiently co-transfected plasmid possessing the firefly luciferase gene under the control of the NPY promoter. This effect of BDNF was completely blocked by either a Y484F mutation in TrkB (which blocks high-affinity Shc binding to TrkB) or by a Y785F substitution [which blocks the binding, phosphorylation and activation of phospholipase Cgamma (PLCgamma)]. Activation of the NPY promoter by neurotrophin-3 in PC12 cells overexpressing TrkC was also completely blocked by a naturally occurring kinase insert which prevents the high-affinity binding of Shc and PLCgamma. NPY promoter activation by BDNF was blocked by PD98059, suggesting a role for mitogen-activated protein kinase (MAP kinase). Stimulation of NPY gene expression by PMA, but not by BDNF, was blocked by Ro-31-8220, a protein kinase C inhibitor, excluding a role for this serine/threonine protein kinase in the effect of BDNF. In addition, BDNF did not cause an elevation in cytosolic Ca2+ concentration. Taken together, our results suggest that stimulation of the NPY promoter by BDNF requires the simultaneous activation of two distinct pathways; one involves Shc and MAP kinase, and the other appears to be PLCgamma-independent but requires an intact tyrosine-785 on TrkB and so may involve an effector of TrkB signalling that remains to be identified.

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Selected References

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  1. Alessi D. R., Cuenda A., Cohen P., Dudley D. T., Saltiel A. R. PD 098059 is a specific inhibitor of the activation of mitogen-activated protein kinase kinase in vitro and in vivo. J Biol Chem. 1995 Nov 17;270(46):27489–27494. doi: 10.1074/jbc.270.46.27489. [DOI] [PubMed] [Google Scholar]
  2. Allen J. M., Martin J. B., Heinrich G. Neuropeptide Y gene expression in PC12 cells and its regulation by nerve growth factor: a model for developmental regulation. Brain Res. 1987 Dec;427(1):39–43. doi: 10.1016/0169-328x(87)90042-8. [DOI] [PubMed] [Google Scholar]
  3. Balbi D., Allen J. M. Role of protein kinase C in mediating NGF effect on neuropeptide Y expression in PC12 cells. Brain Res Mol Brain Res. 1994 Jun;23(4):310–316. doi: 10.1016/0169-328x(94)90240-2. [DOI] [PubMed] [Google Scholar]
  4. Baxter R. M., Cohen P., Obermeier A., Ullrich A., Downes C. P., Doza Y. N. Phosphotyrosine residues in the nerve-growth-factor receptor (Trk-A). Their role in the activation of inositolphospholipid metabolism and protein kinase cascades in phaeochromocytoma (PC12) cells. Eur J Biochem. 1995 Nov 15;234(1):84–91. doi: 10.1111/j.1432-1033.1995.084_c.x. [DOI] [PubMed] [Google Scholar]
  5. Bush A. B., Borden L. A., Greene L. A., Maxfield F. R. Nerve growth factor potentiates bradykinin-induced calcium influx and release in PC12 cells. J Neurochem. 1991 Aug;57(2):562–574. doi: 10.1111/j.1471-4159.1991.tb03787.x. [DOI] [PubMed] [Google Scholar]
  6. Canossa M., Rovelli G., ShooterEM Transphosphorylation of the neurotrophin Trk receptors. J Biol Chem. 1996 Mar 8;271(10):5812–5818. doi: 10.1074/jbc.271.10.5812. [DOI] [PubMed] [Google Scholar]
  7. Damon D. H., D'Amore P. A., Wagner J. A. Nerve growth factor and fibroblast growth factor regulate neurite outgrowth and gene expression in PC12 cells via both protein kinase C- and cAMP-independent mechanisms. J Cell Biol. 1990 Apr;110(4):1333–1339. doi: 10.1083/jcb.110.4.1333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. De Bernardi M. A., Rabins S. J., Colangelo A. M., Brooker G., Mocchetti I. TrkA mediates the nerve growth factor-induced intracellular calcium accumulation. J Biol Chem. 1996 Mar 15;271(11):6092–6098. doi: 10.1074/jbc.271.11.6092. [DOI] [PubMed] [Google Scholar]
  9. Dickens G., Lavarreda M., Zheng W. H., Guroff G. Involvement of protein kinase C in nerve growth factor- and K-252a-stimulated calcium uptake into PC12 cells. J Neurosci Res. 1997 Feb 1;47(3):271–276. doi: 10.1002/(sici)1097-4547(19970201)47:3<271::aid-jnr5>3.0.co;2-f. [DOI] [PubMed] [Google Scholar]
  10. Ginty D. D., Fanger G. R., Wagner J. A., Maue R. A. The activity of cAMP-dependent protein kinase is required at a posttranslational level for induction of voltage-dependent sodium channels by peptide growth factors in PC12 cells. J Cell Biol. 1992 Mar;116(6):1465–1473. doi: 10.1083/jcb.116.6.1465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Greene L. A., Tischler A. S. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2424–2428. doi: 10.1073/pnas.73.7.2424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guiton M., Gunn-Moore F. J., Glass D. J., Geis D. R., Yancopoulos G. D., Tavaré J. M. Naturally occurring tyrosine kinase inserts block high affinity binding of phospholipase C gamma and Shc to TrkC and neurotrophin-3 signaling. J Biol Chem. 1995 Sep 1;270(35):20384–20390. doi: 10.1074/jbc.270.35.20384. [DOI] [PubMed] [Google Scholar]
  13. Guiton M., Gunn-Moore F. J., Stitt T. N., Yancopoulos G. D., Tavaré J. M. Identification of in vivo brain-derived neurotrophic factor-stimulated autophosphorylation sites on the TrkB receptor tyrosine kinase by site-directed mutagenesis. J Biol Chem. 1994 Dec 2;269(48):30370–30377. [PubMed] [Google Scholar]
  14. Gunn-Moore F. J., Williams A. G., Tavaré J. M. Analysis of mitogen-activated protein kinase activation by naturally occurring splice variants of TrkC, the receptor for neurotrophin-3. Biochem J. 1997 Feb 15;322(Pt 1):193–198. doi: 10.1042/bj3220193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hawley R. J., Scheibe R. J., Wagner J. A. NGF induces the expression of the VGF gene through a cAMP response element. J Neurosci. 1992 Jul;12(7):2573–2581. doi: 10.1523/JNEUROSCI.12-07-02573.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Heasley L. E., Johnson G. L. Regulation of protein kinase C by nerve growth factor, epidermal growth factor, and phorbol esters in PC12 pheochromocytoma cells. J Biol Chem. 1989 May 25;264(15):8646–8652. [PubMed] [Google Scholar]
  17. Higuchi H., Nakano K., Miki N. Identification of NGF-response element in the rat neuropeptide Y gene and induction of the binding proteins. Biochem Biophys Res Commun. 1992 Dec 30;189(3):1553–1560. doi: 10.1016/0006-291x(92)90253-h. [DOI] [PubMed] [Google Scholar]
  18. Kalman D., Wong B., Horvai A. E., Cline M. J., O'Lague P. H. Nerve growth factor acts through cAMP-dependent protein kinase to increase the number of sodium channels in PC12 cells. Neuron. 1990 Mar;4(3):355–366. doi: 10.1016/0896-6273(90)90048-k. [DOI] [PubMed] [Google Scholar]
  19. Kobayashi E., Nakano H., Morimoto M., Tamaoki T. Calphostin C (UCN-1028C), a novel microbial compound, is a highly potent and specific inhibitor of protein kinase C. Biochem Biophys Res Commun. 1989 Mar 15;159(2):548–553. doi: 10.1016/0006-291x(89)90028-4. [DOI] [PubMed] [Google Scholar]
  20. Lamballe F., Tapley P., Barbacid M. trkC encodes multiple neurotrophin-3 receptors with distinct biological properties and substrate specificities. EMBO J. 1993 Aug;12(8):3083–3094. doi: 10.1002/j.1460-2075.1993.tb05977.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Machida C. M., Scott J. D., Ciment G. NGF-induction of the metalloproteinase-transin/stromelysin in PC12 cells: involvement of multiple protein kinases. J Cell Biol. 1991 Sep;114(5):1037–1048. doi: 10.1083/jcb.114.5.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Marshall C. J. Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell. 1995 Jan 27;80(2):179–185. doi: 10.1016/0092-8674(95)90401-8. [DOI] [PubMed] [Google Scholar]
  23. Middlemas D. S., Meisenhelder J., Hunter T. Identification of TrkB autophosphorylation sites and evidence that phospholipase C-gamma 1 is a substrate of the TrkB receptor. J Biol Chem. 1994 Feb 18;269(7):5458–5466. [PubMed] [Google Scholar]
  24. Milbrandt J. Nerve growth factor rapidly induces c-fos mRNA in PC12 rat pheochromocytoma cells. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4789–4793. doi: 10.1073/pnas.83.13.4789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Minth-Worby C. A. Transcriptional regulation of the human neuropeptide Y gene by nerve growth factor. J Biol Chem. 1994 Jun 3;269(22):15460–15468. [PubMed] [Google Scholar]
  26. Nikodijevic B., Guroff G. Nerve growth factor-induced increase in calcium uptake by PC12 cells. J Neurosci Res. 1991 Feb;28(2):192–199. doi: 10.1002/jnr.490280206. [DOI] [PubMed] [Google Scholar]
  27. Obermeier A., Bradshaw R. A., Seedorf K., Choidas A., Schlessinger J., Ullrich A. Neuronal differentiation signals are controlled by nerve growth factor receptor/Trk binding sites for SHC and PLC gamma. EMBO J. 1994 Apr 1;13(7):1585–1590. doi: 10.1002/j.1460-2075.1994.tb06421.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Obermeier A., Halfter H., Wiesmüller K. H., Jung G., Schlessinger J., Ullrich A. Tyrosine 785 is a major determinant of Trk--substrate interaction. EMBO J. 1993 Mar;12(3):933–941. doi: 10.1002/j.1460-2075.1993.tb05734.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Obermeier A., Lammers R., Wiesmüller K. H., Jung G., Schlessinger J., Ullrich A. Identification of Trk binding sites for SHC and phosphatidylinositol 3'-kinase and formation of a multimeric signaling complex. J Biol Chem. 1993 Nov 5;268(31):22963–22966. [PubMed] [Google Scholar]
  30. Ohmichi M., Zhu G., Saltiel A. R. Nerve growth factor activates calcium-insensitive protein kinase C-epsilon in PC-12 rat pheochromocytoma cells. Biochem J. 1993 Nov 1;295(Pt 3):767–772. doi: 10.1042/bj2950767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pandiella-Alonso A., Malgaroli A., Vicentini L. M., Meldolesi J. Early rise of cytosolic Ca2+ induced by NGF in PC12 and chromaffin cells. FEBS Lett. 1986 Nov 10;208(1):48–51. doi: 10.1016/0014-5793(86)81529-0. [DOI] [PubMed] [Google Scholar]
  32. Sabol S. L., Higuchi H. Transcriptional regulation of the neuropeptide Y gene by nerve growth factor: antagonism by glucocorticoids and potentiation by adenosine 3',5'-monophosphate and phorbol ester. Mol Endocrinol. 1990 Mar;4(3):384–392. doi: 10.1210/mend-4-3-384. [DOI] [PubMed] [Google Scholar]
  33. Schwartz M. W., Seeley R. J., Campfield L. A., Burn P., Baskin D. G. Identification of targets of leptin action in rat hypothalamus. J Clin Invest. 1996 Sep 1;98(5):1101–1106. doi: 10.1172/JCI118891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Stephens R. M., Loeb D. M., Copeland T. D., Pawson T., Greene L. A., Kaplan D. R. Trk receptors use redundant signal transduction pathways involving SHC and PLC-gamma 1 to mediate NGF responses. Neuron. 1994 Mar;12(3):691–705. doi: 10.1016/0896-6273(94)90223-2. [DOI] [PubMed] [Google Scholar]
  35. Tatemoto K., Carlquist M., Mutt V. Neuropeptide Y--a novel brain peptide with structural similarities to peptide YY and pancreatic polypeptide. Nature. 1982 Apr 15;296(5858):659–660. doi: 10.1038/296659a0. [DOI] [PubMed] [Google Scholar]
  36. Traverse S., Gomez N., Paterson H., Marshall C., Cohen P. Sustained activation of the mitogen-activated protein (MAP) kinase cascade may be required for differentiation of PC12 cells. Comparison of the effects of nerve growth factor and epidermal growth factor. Biochem J. 1992 Dec 1;288(Pt 2):351–355. doi: 10.1042/bj2880351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Treisman R. Ternary complex factors: growth factor regulated transcriptional activators. Curr Opin Genet Dev. 1994 Feb;4(1):96–101. doi: 10.1016/0959-437x(94)90097-3. [DOI] [PubMed] [Google Scholar]
  38. Valenzuela D. M., Maisonpierre P. C., Glass D. J., Rojas E., Nuñez L., Kong Y., Gies D. R., Stitt T. N., Ip N. Y., Yancopoulos G. D. Alternative forms of rat TrkC with different functional capabilities. Neuron. 1993 May;10(5):963–974. doi: 10.1016/0896-6273(93)90211-9. [DOI] [PubMed] [Google Scholar]
  39. Young S. W., Dickens M., Tavaré J. M. Differentiation of PC12 cells in response to a cAMP analogue is accompanied by sustained activation of mitogen-activated protein kinase. Comparison with the effects of insulin, growth factors and phorbol esters. FEBS Lett. 1994 Jan 31;338(2):212–216. doi: 10.1016/0014-5793(94)80367-6. [DOI] [PubMed] [Google Scholar]
  40. Zirrgiebel U., Ohga Y., Carter B., Berninger B., Inagaki N., Thoenen H., Lindholm D. Characterization of TrkB receptor-mediated signaling pathways in rat cerebellar granule neurons: involvement of protein kinase C in neuronal survival. J Neurochem. 1995 Nov;65(5):2241–2250. doi: 10.1046/j.1471-4159.1995.65052241.x. [DOI] [PubMed] [Google Scholar]

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