TrkB Regulates N-Methyl-D-Aspartate Receptor Signaling by Uncoupling and Recruiting the Brain-Specific Guanine Nucleotide Exchange Factor, RasGrf1

  • Asghar Talebian
  • Kim Robinson-Brookes
  • Susan O. Meakin


Brain-derived neurotrophic factor (BDNF) facilitates multiple aspects of neuronal differentiation and cellular physiology by activating the high-affinity receptor tyrosine kinase, TrkB. While it is known that both BDNF and TrkB modulate cellular processes involved in learning and memory, exactly how TrkB cross-talks and modulates signaling downstream of excitatory ionotropic receptors, such as the NMDA receptor (NMDAR), are not well understood. A model that we have investigated involves the signaling molecule RasGrf1, a guanine nucleotide exchange factor for both Ras and Rac. We previously identified RasGrf1 as a novel Trk binding partner that facilitates neurite outgrowth in response to both nerve growth factor (NGF) (Robinson et al. in J Biol Chem 280:225–235, 2005) and BDNF (Talebian et al. in J Mol Neurosci 49:38–51, 2013); however, RasGrf1 can also bind the NR2B subunit of the NMDAR (Krapivinsky et al. in Neuron 40:775–784, 2003) and stimulate long-term depression (LTD) (Li et al. in J Neurosci 26:1721–1729, 2006). We have addressed a model that TrkB facilitates learning and memory via two processes. First, TrkB uncouples RasGrf1 from NR2B and facilitates a decrease in NMDA signaling associated with LTD (p38-MAPK). Second, the recruitment of RasGrf1 to TrkB enhances neurite outgrowth and pERK activation and signaling associated with learning and memory. We demonstrate that NMDA recruits RasGrf1 to NR2B; however, co-stimulation with BDNF uncouples this association and recruits RasGrf1 to TrkB. In addition, activation of TrkB stimulates the tyrosine phosphorylation of RasGrf1 which increases neurite outgrowth (Talebian et al. in J Mol Neurosci 49:38–51, 2013), and the tyrosine phosphorylation of NR2B (Tyr1472) (Nakazawa et al. in J Biol Chem 276:693–699, 2001) which facilitates NMDAR cell surface retention (Zhang et al. in J Neurosci 28:415–24, 2008). Collectively, these data demonstrate that TrkB alters NMDA signaling by a dual mechanism that uncouples LTD and, in turn, stimulates neuronal growth and the signaling pathways associated with learning and memory.


N-methyl-D-aspartate (NMDA) receptor Ras guanine-nucleotide exchange factor (RasGrf1) Trophomyosin-related kinase B (TrkB) Phosphotyrosine signaling Neurobiology Brain-derived neurotrophic factor (BDNF) 


Author Contribution

SOM conceived the study. KRB and AT conducted the experiments and all three authors contributed to writing the paper.

Funding Information

This work was supported by a Discovery Grant to Susan O. Meakin from the Natural Sciences and Engineering Research Council of Canada (RGPIN 341947). K. Robinson-Brookes and Asghar Talebian were supported with funds from an Ontario Graduate Scholarship and an Ontario Graduate Scholarship in Science and Technology respectively.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. Agamasu C, Ghanam RH, Saad JS (2015) Structural and biophysical characterization of the interactions between calmodulin and the pleckstrin homology domain of Akt. J Biol Chem 290:27403–27413CrossRefPubMedCentralGoogle Scholar
  2. Anborgh PH, Qian X, Papageorge AG, Vass WC, DeClue JE, Lowy DR (1999) Ras-specific exchange factor GRF: oligomerization through its Dbl homology domain and calcium-dependent activation of Raf. Mol Cell Biol 19:4611–4622CrossRefPubMedCentralGoogle Scholar
  3. Atwal JK, Massie B, Miller FD, Kaplan DR (2000) The TrkB-Shc site signals neuronal survival and local axon growth via MEK and PI3-kinase. Neuron 27:265–277CrossRefPubMedCentralGoogle Scholar
  4. Baouz S, Jacquet E, Accorsi K, Hountondji C, Balestrini M, Zippel R, Sturani E, Parmeggiani A (2001) Sites of phosphorylation by protein kinase A in CDC25Mm/GRF1, a guanine nucleotide exchange factor for Ras. J Biol Chem 276:1742–1749CrossRefPubMedCentralGoogle Scholar
  5. Carvalho A (2008) Role of the brain-derived neurotrophic factor at glutamatergic synapses. Br J Pharmacol 153:310–324CrossRefGoogle Scholar
  6. Chapleau CA, Carlo ME, Larimore JL, Pozzo-Miller L (2008) The actions of BDNF on dendritic spine density and morphology in organotypic slice cultures depend on the presence of serum in culture media. J Neurosci Methods 169:182–190CrossRefPubMedCentralGoogle Scholar
  7. Dell’Acqua L (2006) Regulation of neuronal PKA signaling through AKAP targeting dynamics. Eur J Cell Biol 85:627–633CrossRefPubMedCentralGoogle Scholar
  8. Dixon SJ, MacDonald JIS, Robinson KN, Kubu CJ, Meakin SO (2006) Trk receptor binding and neurotrophin/fibroblast growth factor (FGF)-dependent activation of the FGF receptor substrate (FRS)-3. BBA Mol Cell Res 1763:366–380Google Scholar
  9. Goebel-Goody SM, Davies KD, Alvestad Linger RM, Freund RK, Browning MD (2009) Phosphoregulation of synaptic and extrasynaptic N-methyl-d-aspartate receptors in adult hippocampal slices. Neuroscience 158:1446–1459CrossRefPubMedCentralGoogle Scholar
  10. Gorski JA, Zeiler SR, Tamowski S, Jones KR (2003) Brain-derived neurotrophic factor is required for the maintenance of cortical dendrites. J Neurosci 23:6856–6865CrossRefPubMedCentralGoogle Scholar
  11. Graham SJ, Russell WC, Nairn R (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36:59–74CrossRefPubMedCentralGoogle Scholar
  12. Jang S-W, Liu X, Yepes M, Shepherd KR, Miller GW, Liu Y, Wilson WD, Xiao G, Blanchi B, Sun YE, Ye K (2010) A selective TrkB agonist with potent neurotrophic activities by 7,8-dihydroxyflavone. Proc Natl Acad Sci 107:2687–2692CrossRefPubMedCentralGoogle Scholar
  13. Ji Y, Pang PT, Feng L, Lu B (2005) Cyclic AMP controls BDNF-induced TrkB phosphorylation and dendritic spine formation in mature hippocampal neurons. Nat Neurosci 8:164–172CrossRefPubMedCentralGoogle Scholar
  14. Kakegawa W (2004) Input- and subunit-specific AMPA receptor trafficking underlying long-term potentiation at hippocampal CA3 synapses. Eur J Neurosci 20:101–110CrossRefPubMedCentralGoogle Scholar
  15. Kiyono M, Kato J, Kataoka T, Kaziro Y, Satoh T (2000a) Stimulation of Ras guanine nucleotide exchange activity of Ras-GRF1/CDC25Mm upon tyrosine phosphorylation by the Cdc42-regulated kinase ACK1. J Biol Chem 275:29788–29793CrossRefPubMedCentralGoogle Scholar
  16. Kiyono M, Kaziro Y, Satoh T (2000b) Induction of Rac-guanine nucleotide exchange activity of Ras-GRF1/CDC25Mm following phosphorylation by the nonreceptor tyrosine kinase Src. J Biol Chem 275:5441–5446CrossRefPubMedCentralGoogle Scholar
  17. Kollen M, Dutar P, Jouvenceau A (2008) The magnitude of hippocampal long term depression depends on the synaptic location of activated NR2-containing N-methyl-d-aspartate receptors. Neuroscience 154:1308–1317CrossRefPubMedCentralGoogle Scholar
  18. Krapivinsky G, Krapivinisky L, Manasian Y, Ivanov A, Tyzio R, Pellegrino C, Ben-Ari Y, Clapham DE, Medina I (2003) The NMDA receptor is coupled to the ERK pathway by a direct interaction between NR2B and RasGrf1. Neuron 40:775–784CrossRefPubMedCentralGoogle Scholar
  19. Kutsuwada T (1992) Molecular diversity of the NMDA receptor channel. Nature 358:36–41CrossRefPubMedCentralGoogle Scholar
  20. Lemmon MA (2007) Pleckstrin homology (PH) domains and phosphoinositides. Biochem Soc Symp 74:81–93CrossRefGoogle Scholar
  21. Lemmon M, Ferguson K, Schlessinger J (1996) PH domains: diverse sequences with a common fold recruit signaling molecules to the cell surface. Cell 85:621–624CrossRefPubMedCentralGoogle Scholar
  22. Levine ES, Kolb JE (2000) Brain-derived neurotrophic factor increases activity of the NR2B-containing N-methyl-D-aspartate receptors in excised patches from hippocampal neurons. J Neurosci Res 62:357–363CrossRefPubMedCentralGoogle Scholar
  23. Levine ES, Crozier RA, Black IB, Plummer MR (1998) Brain-derived neurotrophic factor modulates hippocampal synaptic transmission by increasing N-methyl-D-aspartic acid receptor activity. Proc Natl Acad Sci U S A 95:10235–10239CrossRefPubMedCentralGoogle Scholar
  24. Li S, Tian X, Hartley DM, Feig LA (2006) Distinct roles for Ras-guanine nucleotide-releasing factor 1 (RasGrf1) and Ras-Grf2 in the induction of long-term potentiation and long-term depression. J Neurosci 26:1721–1729CrossRefPubMedCentralGoogle Scholar
  25. Lin S-Y, Wu K, Levine ES, Mount HTJ, Suen P-C, Black IB (1998) BDNF acutely increases tyrosine phosphorylation of the NMDA receptor subunit 2B in cortical and hippocampal postsynaptic densities. Mol Brain Res 55:20–27CrossRefPubMedCentralGoogle Scholar
  26. Lisman J (2003) Long-term potentiation: outstanding questions and attempted synthesis. Philos Trans R Soc Lond Ser B Biol Sci 358:829–842CrossRefGoogle Scholar
  27. Liu H-Y, Meakin SO (2002) ShcB and ShcC activation by the Trk family of receptor tyrosine kinases. J Biol Chem 277:26046–26056CrossRefPubMedCentralGoogle Scholar
  28. Liu XB, Murray KD, Jones EG (2004) Switching of NMDA receptor 2A and 2B subunits at thalamic and cortical synapses during early postnatal development. J Neurosci 24:8885–8895CrossRefPubMedCentralGoogle Scholar
  29. Loeb DM, Stephens RM, Copeland T, Kaplan DR, Greene LA (1994) A Trk NGF receptor point mutation affecting interaction with PLCγ-1 abolishes NGF- promoted peripherin induction, but not neurite outgrowth. J Biol Chem 269:8901–8910PubMedPubMedCentralGoogle Scholar
  30. Lu Y, Christian K, Lu B (2008) BDNF: a key regulator for protein synthesis- dependent LTP and long-term memory? Neurobiol Learn Mem 89:312–323CrossRefPubMedCentralGoogle Scholar
  31. MacDonald J (2006) Hippocampal long-term synaptic plasticity and signal amplification of NMDA receptors. Crit Rev Neurobiol 18:71–84CrossRefPubMedCentralGoogle Scholar
  32. MacDonald JIS, Verdi JM, Meakin SO (1999) Activity-dependent interaction of the intracellular domain of rat TrkA with intermediate filament proteins, the β-6 proteasomal subunit, Ras GRF1 and the p162 subunit of eIF3. J Mol Neurosci 13:141–158CrossRefPubMedCentralGoogle Scholar
  33. Manabe T (2002) Does BDNF have pre-or postsynaptic targets? Science 295:1651–1653CrossRefPubMedCentralGoogle Scholar
  34. Massey P (2004) Differential roles of NR2A and NR2B-containing NMDA receptors in cortical long-term potentiation and long-term depression. J Neurosci 24:7821–7828CrossRefPubMedCentralGoogle Scholar
  35. Meakin SO, MacDonald JIS (1998) A novel juxtamembrane deletion in rat TrkA blocks differentiative but not Mitogenic cell Signalling in response to nerve growth factor. J Neurochem 71:1875–1888CrossRefPubMedCentralGoogle Scholar
  36. Meakin SO, MacDonald JIS, Gryz EA, Kubu CJ, Verdi JM (1999) The signaling adapter protein FRS-2 competes with Shc for binding to TrkA: a model for discriminating proliferation and differentiation. J Biol Chem 274:9861–9870CrossRefPubMedCentralGoogle Scholar
  37. Minichiello L, Korte M, Wolfer D, Kühn R, Unsicker K, Cestari V, Rossi- Arnaurd C, Lipp H-P, Bonhoeffer T, Klein R (1999) Essential role for TrkB receptors in hippocampal-mediated learning. Neuron 24:401–414CrossRefPubMedCentralGoogle Scholar
  38. Monyer H (1992) Heteromeric NMDA receptors: molecular and functional distinction of subtypes. Science 256:1217–1221CrossRefPubMedCentralGoogle Scholar
  39. Murphy JA, Stein IS, Lau CG, Peixoto RT, Aman TK, Kaneko N, Aromolaran K, Saulnier JL, Popescu GK, Sabatini BL, Hell JW, Zukin RS (2014) Phosphorylation of Ser1166 on GluN2B by PKA is critical to synaptic NMDA receptor function and Ca2+ signaling in spines. J Neurosci 34:869–879CrossRefPubMedCentralGoogle Scholar
  40. Nakazawa T, Komai S, Tezuka T, Hisatsune C, Umemori H, Semba K, Mishina M, Manabe T, Yamamoto T (2001) Characterization of Fyn-mediated tyrosine phosphorylation sites on GluR ε2 (NR2B) subunit of the N-methyl-D-aspartate receptor. J Biol Chem 276:693–699CrossRefGoogle Scholar
  41. Obermeier A, Bradshaw RA, Seedorf K, Choidas A, Schlessinger J, Ullrich A (1994) Neuronal differentiation signals are controlled by nerve growth factor receptor/Trk binding sites for SHC and PLCγ. EMBO J 13:1585–1590CrossRefPubMedCentralGoogle Scholar
  42. Pawson T, Nash P (2000) Protein-protein interactions define specificity in signal transduction. Genes Dev 14:1027–1047PubMedPubMedCentralGoogle Scholar
  43. Poo MM (2001) Neurotrophins as synaptic modulators. Nat Rev Neurosci 2:24–32CrossRefPubMedCentralGoogle Scholar
  44. Pozzo-Miller L (1999) Impairments in high-frequency transmission, synaptic vesicle docking, and synaptic protein distribution in the hippocampus of BDNF knockout mice. J Neurosci 19:4972–4983CrossRefPubMedCentralGoogle Scholar
  45. Prybylowski K, Chang K, Sans N, Kan L, Vicini S, Wenthold RJ (2005) The synaptic localization of NR2B-containing NMDA receptors is controlled by interactions with PDZ proteins and AP-2. Neuron 47:845–857CrossRefPubMedCentralGoogle Scholar
  46. Robinson KN, Manto K, Buchsbaum RJ, MacDonald JIS, Meakin SO (2005) Neurotrophin-dependent tyrosine phosphorylation of Ras guanine-releasing factor 1 and associated neurite outgrowth is dependent on the HIKE domain of TrkA. J Biol Chem 280:225–235CrossRefPubMedCentralGoogle Scholar
  47. Rose CR, Blum R, Kafitz KW, Kovalchuk Y, Konnerth A (2004) From modulator to mediator: rapid effects of BDNF on ion channels. BioEssays 26:1185–1194CrossRefPubMedCentralGoogle Scholar
  48. Schinder AF, Poo M (2000) The neurotrophin hypothesis for synaptic plasticity. Trends Neurosci 23:639–645CrossRefPubMedCentralGoogle Scholar
  49. Skeberdis VA, Chevaleyre V, Lau CG, Goldberg JH, Pettit DL, Suadicani SO, Lin Y, Bennett MVL, Yuste R, Castillo PE, Zukin RS (2006) Protein kinase A regulates calcium permeability of NMDA receptors. Nat Neurosci 9:501–510CrossRefPubMedCentralGoogle Scholar
  50. Stephens RM, Loeb DM, Copeland TD, Pawson T, Greene LA, Kaplan DR (1994) Trk receptors use redundant signal transduction pathways involving SHC and PLC- γ1 to mediate NGF responses. Neuron 12:691–705CrossRefPubMedCentralGoogle Scholar
  51. Sturani E, Abbondio A, Branduardi P, Ferrari C, Zippel R, Martegani E, Vanoni M, Denis-Donini S (1997) The ras guanine nucleotide exchange factor cdc25Mm is present at the synaptic junction. Exp Cell Res 235:117–123CrossRefPubMedCentralGoogle Scholar
  52. Talebian A, Robinson-Brookes K, MacDonald JIS, Meakin SO (2013) Ras guanine nucleotide releasing factor 1 (RasGrf1) enhancement of Trk receptor mediated neurite outgrowth requires activation of both H-Ras and Rac. J Mol Neurosci 49:38–51CrossRefPubMedCentralGoogle Scholar
  53. Tanaka J (2008) Protein synthesis and neurotrophin-dependent structural plasticity of single dendritic spines. Science 319:1683–1687CrossRefPubMedCentralGoogle Scholar
  54. Taniguchi N (2006) Possible involvement of BDNF release in long-lasting synapse formation induced by repetitive PKA activation. Neurosci Lett 406:38–42CrossRefPubMedCentralGoogle Scholar
  55. Tian GT, Tsuji K, Lo EH, Huang S, Feig LA (2004) Developmentally regulated role for Ras-GRFs in coupling NMDA glutamate receptors to Ras, Erk and CREB. EMBO J 23:1567–1575CrossRefPubMedCentralGoogle Scholar
  56. van Zundert B, Yoshii A, Constantine-Paton M (2004) Receptor compartmentalization and trafficking at glutamate synapses: a developmental proposal. Trends Neurosci 27:428–437CrossRefPubMedCentralGoogle Scholar
  57. Wu H-Y, Hsu F-C, Gleichman AJ, Baconguis I, Coulter DA, Lynch DR (2007) Fyn-mediated phosphorylation of NR2B Tyr-1336 controls calpain-mediated NR2B cleavage in neurons and heterologous systems. J Biol Chem 282:20075–20087CrossRefPubMedCentralGoogle Scholar
  58. Xu B, Gottschalk W, Chow A, Wilson RI, Schnell E, Zang K, Wang D, Nicoll RA, Lu B, Reichardt LF (2000) The role of brain-derived neurotrophic factor receptors in the mature Hippocamus: modulation of long-term potentiation through a presynaptic mechanism involving TrkB. J Neurosci 20:6888–6897CrossRefPubMedCentralGoogle Scholar
  59. Yamada K (2003) Brain-derived neurotrophic factor/TrkB signaling in memory processes. J Pharmacol Sci 91:267–270CrossRefPubMedCentralGoogle Scholar
  60. Yamada K (2004) Interaction of BDNF/TrkB signaling with NMDA receptor in learning and memory. Drug News Perspect 17:435–438CrossRefPubMedCentralGoogle Scholar
  61. Zhang S, Edelmann L, Liu J, Crandall JE, Morabito MA (2008) Cdk5 regulates the phosphorylation of Tyrosine 1472 NR2B and the surface expression of NMDA receptors. J Neurosci 28:415–424CrossRefPubMedCentralGoogle Scholar
  62. Zhang Z, Fan J, Ren Y, Zhou W, Yin G (2013) The release of glutamate from cortical neurons regulated by BDNF via the TrkB/Src/PLC-γ1 pathway. J Cell Biochem 114:144–151CrossRefPubMedCentralGoogle Scholar
  63. Zippel R, Gnesutta N, Matus-Leibovitch N, Mancinelli E, Saya D, Vogel Z, Sturani E, Renata Z, Nerina G, Noa ML, Enzo M, Daniella S, Zvi V, Emmapaola S (1997) Ras-Grf, the activator of Ras, is expressed preferentially in mature neurons of the central nervous system. Brain Res Mol Brain Res 48:140–144CrossRefPubMedCentralGoogle Scholar

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Authors and Affiliations

  1. 1.Department of BiochemistryWestern UniversityLondonCanada
  2. 2.Graduate Program in Neuroscience, Western UniversityLondonCanada

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