Journal of Neural Transmission

, Volume 124, Issue 6, pp 749–759 | Cite as

The hallucinogen 2,5-dimethoxy-4-iodoamphetamine hydrochloride activates neurotrophin receptors in a neuronal cell line and promotes neurites extension

Translational Neurosciences - Original Article


Decreased neurotrophic factors expression and neurotrophin receptors signalling have repeatedly been reported in association with stress, depression, and neurodegenerative disorders. We have previously identified the hallucinogen 2,5-dimethoxy-4-iodoamphetamine hydrochloride (DOI) as protective against trophic deprivation-induced cytotoxicity in human neuroblastoma SK-N-SH cells and established the dependence of this effect on the 5-HT2A receptor, tyrosine kinases activity, and the extracellular signal-regulated kinase pathway. In the current study, we investigated the effect of DOI on tropomyosin-related kinase receptor A (TrkA) phosphorylation. Treatment with DOI increased TrkA tyrosine phosphorylation in SK-N-SH cells, determined by immunoprecipitation with TrkA antibody and immunoblotting with anti-phosphotyrosine- and TrkA-antibodies. Analysis of DOI’s effect on individual TrkA residues in SK-N-SH cells showed that it increases TrkA Tyr490 phosphorylation (177 ± 23% after 5 μM DOI for 30 min compared to vehicle). Furthermore, DOI treatment increased the percentage of SK-N-SH cells extending neurites in a TrkA-dependent manner (17.2 ± 2.2% after 5 μM DOI treatment for 6 days compared to 5.6 ± 1.7% after vehicle). In a different cell model—lymphoblastoid cell lines—DOI treatment increased tropomyosin-related kinase receptor B (TrkB) phosphorylation, determined by immunoprecipitation with TrkB antibody and immunoblotting with anti-phosphotyrosine antibody and total Trk antibody. Our results identify the Trk receptors as a downstream target of the hallucinogen DOI. In light of the known involvement of Trk receptors in mental diseases, their participation in DOI-mediated effects warrants further investigation.


Hallucinogen Tropomyosin-related kinase receptors SK-N-SH neuroblastoma cells Neurites 



Serotonin 2


Tropomyosin-related kinase receptor A


Tropomyosin-related kinase receptor B


2,5-Dimethoxy-4-iodoamphetamine hydrochloride


Brain-derived neurotrophic factor


Extracellular signal-regulated kinase


Nerve growth factor






Mitogen-activated protein kinase


Phosphatidylinositol 3-kinase


Protein kinase B


Phospholipase C-γ


Protein kinase C


Fetal bovine serum


Phosphate-buffered saline


Epstein-Barr virus




Analysis of variance


G protein-coupled receptors


Least statistical difference



Financial support for the study was provided through the Marie Heim-Vögtlin program of the Swiss National Science Foundation.

Supplementary material

702_2017_1706_MOESM1_ESM.doc (1.4 mb)
Supplementary material 1 (DOC 1411 kb)


  1. Aloe L, Rocco ML, Bianchi P, Manni L (2012) Nerve growth factor: from the early discoveries to the potential clinical use. J Transl Med 10:239CrossRefPubMedPubMedCentralGoogle Scholar
  2. Arévalo JC, Wu SH (2006) Neurotrophin signaling: many exciting surprises! Cell Mol Life Sci 63:1523–1537CrossRefPubMedGoogle Scholar
  3. Baxter RM, Cohen P, Obermeier A, Ullrich A, Downes CP, Doza YN (1995) 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 234:84–91CrossRefPubMedGoogle Scholar
  4. Bhatia HS, Agrawal R, Sharma S, Huo YX, Ying Z, Gomez-Pinilla F (2011) Omega-3 fatty acid deficiency during brain maturation reduces neuronal and behavioral plasticity in adulthood. PLoS One 6:e28451CrossRefPubMedPubMedCentralGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  6. Dudok JJ, Groffen AJ, Witter MP, Voorn P, Verhage M (2009) Chronic activation of the 5-HT(2) receptor reduces 5-HT neurite density as studied in organotypic slice cultures. Brain Res 1302:1–9CrossRefPubMedGoogle Scholar
  7. Hartman DS, McCormack M, Schubenel R, Hertel C (1992) Multiple trkA proteins in PC12 cells bind NGF with a slow association rate. J Biol Chem 267:24516–24522PubMedGoogle Scholar
  8. Hellemans J, Mortier G, De Paepe A, Speleman F, Vandesompele J (2007) qBase relative quantification framework and software for management and automated analysis of real-time quantitative PCR data. Genome Biol 8:R19CrossRefPubMedPubMedCentralGoogle Scholar
  9. Hwang JJ, Park MH, Choi SY, Koh JY (2005) Activation of the Trk signaling pathway by extracellular zinc. Role of metalloproteinases. J Biol Chem 280:11995–12001CrossRefPubMedGoogle Scholar
  10. Jang SW, Liu X, Chan CB, Weinshenker D, Hall RA, Xiao G, Ye K (2009) Amitriptyline is a TrkA and TrkB receptor agonist that promotes TrkA/TrkB heterodimerization and has potent neurotrophic activity. Chem Biol 16:644–656CrossRefPubMedPubMedCentralGoogle Scholar
  11. Jang SW, Liu X, Pradoldej S, Tosini G, Chang Q, Iuvone PM, Ye K (2010) N-acetylserotonin activates TrkB receptor in a circadian rhythm. Proc Natl Acad Sci USA 107:3876–3881CrossRefPubMedPubMedCentralGoogle Scholar
  12. Klongpanichapak S, Phansuwan-Pujito P, Ebadi M, Govitrapong P (2008) Melatonin inhibits amphetamine-induced increase in alpha-synuclein and decrease in phosphorylated tyrosine hydroxylase in SK-N-SH cells. Neurosci Lett 436:309–313CrossRefPubMedGoogle Scholar
  13. Kruk JS, Vasefi MS, Heikkila JJ, Beazely MA (2013) Reactive oxygen species are required for 5-HT-induced transactivation of neuronal platelet-derived growth factor and TrkB receptors, but not for ERK1/2 activation. PLoS One 8:e77027CrossRefPubMedPubMedCentralGoogle Scholar
  14. Lee FS, Chao MV (2001) Activation of Trk neurotrophin receptors in the absence of neurotrphins. Proc Natl Acad Sci 98:3555–3560CrossRefPubMedPubMedCentralGoogle Scholar
  15. Lee FS, Rajagopal R, Kim AH, Chang PC, Chao MV (2002) Activation of Trk neurotrophin receptor signaling by pituitary adenylate cyclase-activating polypeptides. J Biol Chem 277:9096–9102CrossRefPubMedGoogle Scholar
  16. Linker R, Gold R, Luhder F (2009) Function of neurotrophic factors beyond the nervous system: inflammation and autoimmune demyelination. Crit Rev Immunol 29:43–68CrossRefPubMedGoogle Scholar
  17. Marinova Z, Walitza S, Grünblatt E (2013) 5-HT2A serotonin receptor agonist DOI alleviates cytotoxicity in neuroblastoma cells: role of the ERK pathway. Prog Neuropsychopharmacol Biol Psychiatry 44:64–72CrossRefPubMedGoogle Scholar
  18. Meller R, Babity JM, Grahame-Smith DG (2002) 5-HT2A receptor activation leads to increased BDNF mRNA expression in C6 glioma cells. Neuromolecular Med. 1:197–205CrossRefPubMedGoogle Scholar
  19. Nichols DE (2004) Hallucinogens. Pharmacol Ther 10:131–181CrossRefGoogle Scholar
  20. Ohtani A, Kozono N, Senzaki K, Shiga T (2014) Serotonin 2A receptor regulates microtubule assembly and induces dynamics of dendritic growth cones in rat cortical neurons in vitro. Neurosci Res 81–82:11–12CrossRefPubMedGoogle Scholar
  21. Patapoutian A, Reichardt LF (2001) Trk receptors: mediators of neurotrophin action. Curr Opin Neurobiol 11:272–280CrossRefPubMedGoogle Scholar
  22. Persico AM, Di Pino G, Levitt P (2006) Multiple receptors mediate the trophic effects of serotonin on ventroposterior thalamic neurons in vitro. Brain Res 1095:17–25CrossRefPubMedGoogle Scholar
  23. Piiper A, Dikic I, Lutz MP, Leser J, Kronenberger B, Elez R, Cramer H, Müller-Esterl W, Zeuzem S (2002) Cyclic AMP induces transactivation of the receptors for epidermal growth factor and nerve growth factor, thereby modulating activation of MAP kinase, Akt, and neurite outgrowth in PC12 cells. J Biol Chem 277:43623–43630CrossRefPubMedGoogle Scholar
  24. Porter RH, Benwell KR, Lamb H, Malcolm CS, Allen NH, Revell DF, Adams DR, Sheardown MJ (1999) Functional characterization of agonists at recombinant human 5-HT2A, 5-HT2B and 5-HT2C receptors in CHO-K1 cells. Br J Pharmacol 128:13–20CrossRefPubMedPubMedCentralGoogle Scholar
  25. Ramakers C, Ruijter JM, Deprez RH, Moorman AF (2003) Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data. Neurosci Lett 339:62–66CrossRefPubMedGoogle Scholar
  26. Reichardt LF (2006) Neurotrophin-regulated signalling pathways. Philos Trans R Soc Lond B Biol Sci 361:1545–1564CrossRefPubMedPubMedCentralGoogle Scholar
  27. Schulte-Herbrüggen O, Braun A, Rochlitzer S, Jockers-Scherübl MC, Hellweg R (2007) Neurotrophic factors–a tool for therapeutic strategies in neurological, neuropsychiatric and neuroimmunological diseases? Curr Med Chem 14:2318–2329CrossRefPubMedGoogle Scholar
  28. Segal RA, Bhattacharyya A, Rua LA, Alberta JA, Stephens RM, Kaplan DR, Stiles CD (1996) Differential utilization of Trk autophosphorylation sites. J Biol Chem 271:20175–20181CrossRefPubMedGoogle Scholar
  29. Seitz G, Gebhardt S, Beck JF, Böhm W, Lode HN, Niethammer D, Bruchelt G (1998) Ascorbic acid stimulates DOPA synthesis and tyrosine hydroxylase gene expression in the human neuroblastoma cell line SK-N-SH. Neurosci Lett 244:33–36CrossRefPubMedGoogle Scholar
  30. Shen J, Maruyama IN (2011) Nerve growth factor receptor TrkA exists as a preformed, yet inactive, dimer in living cells. FEBS Lett 585:295–299CrossRefPubMedGoogle Scholar
  31. Shen J, Ghai K, Sompol P, Liu X, Cao X, Iuvone PM, Ye K (2012) N-acetyl serotonin derivatives as potent neuroprotectants for retinas. Proc Natl Acad Sci USA 109:3540–3545CrossRefPubMedPubMedCentralGoogle Scholar
  32. Sobreviela T, Clary DO, Reichardt LF, Brandabur MM, Kordower JH, Mufson EJ (1994) TrkA-immunoreactive profiles in the central nervous system: colocalization with neurons containing p75 nerve growth factor receptor, choline acetyltransferase, and serotonin. J Comp Neurol 350:587–611CrossRefPubMedPubMedCentralGoogle Scholar
  33. Stephens RM, Loeb DM, Copeland TD, Pawson T, Greene LA, Kaplan DR (1994) Trk receptors use redundant signal transduction pathways involving SHC and PLC-gamma 1 to mediate NGF responses. Neuron 12:691–705CrossRefPubMedGoogle Scholar
  34. Toyohira Y, Ueno S, Tsutsui M, Itoh H, Sakai N, Saito N, Takahashi K, Yanagihara N (2010) Stimulatory effects of the soy phytoestrogen genistein on noradrenaline transporter and serotonin transporter activity. Mol Nutr Food Res 54:516–524CrossRefPubMedGoogle Scholar
  35. Tsuchioka M, Takebayashi M, Hisaoka K, Maeda N, Nakata Y (2008) Serotonin (5-HT) induces glial cell line-derived neurotrophic factor (GDNF) mRNA expression via the transactivation of fibroblast growth factor receptor 2 (FGFR2) in rat C6 glioma cells. J Neurochem 106:244–257CrossRefPubMedGoogle Scholar
  36. Vega JA, García-Suárez O, Hannestad J, Pérez-Pérez M, Germanà A (2003) Neurotrophins and the immune system. J Anat 203:1–19CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Wien 2017

Authors and Affiliations

  1. 1.Department of Child and Adolescent Psychiatry and Psychotherapy, Psychiatric HospitalUniversity of ZürichZurichSwitzerland
  2. 2.Neuroscience Center ZürichUniversity of Zürich and ETH ZürichZurichSwitzerland
  3. 3.Zürich Center for Integrative Human PhysiologyUniversity of ZürichZurichSwitzerland
  4. 4.Department of Psychosomatic MedicineClinic BarmelweidBarmelweidSwitzerland

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