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Cited by 92Neurotransmitter Receptor Influence on BehaviorReceptor Mechanisms and SignalingNeuroscience and Neuropharmacology Research

Amphetamines signal through intracellular TAAR1 receptors coupled to Gα13 and GαS in discrete subcellular domains

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Underhill, Suzanne M., Hullihen, Patrick D., Chen, Jingshan, Fenollar-Ferrer, Cristina, Rizzo, M. A., Ingram, Susan L., Amara, Susan G.·2021·Molecular Psychiatry
DOI 10.1038/s41380-019-0469-2PMC PMC7038576
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Abstract

The extensive use of amphetamines to treat attention deficit hyperactivity disorders in children provides a compelling rationale for understanding the mechanisms of action of amphetamines and amphetamine-related drugs. We have previously shown that acute amphetamine (AMPH) regulates the trafficking of both dopamine and glutamate transporters in dopamine neurons by increasing activation of the small GTPase RhoA and of protein kinase A. Here we demonstrate that these downstream signaling events depend upon the direct activation of a trace amine-associated receptor, TAAR1, an intracellular G-protein coupled receptor (GPCR) that can be activated by amphetamines, trace amines, and biogenic amine metabolites. Using cell lines and mouse lines in which TAAR1 expression has been disrupted, we demonstrate that TAAR1 mediates the effects of AMPH on both RhoA and cAMP signaling. Inhibition of different Gα signaling pathways in cell lines and in vivo using small cell-permeable peptides confirms that the endogenous intracellular TAAR1 couples to G 13 and to G S α-subunits to increase RhoA and PKA activity, respectively. Results from experiments with RhoA- and PKA-FRET sensors targeted to different subcellular compartments indicate that AMPH-elicited PKA activation occurs throughout the cell, whereas G 13-mediated RhoA activation is concentrated near the endoplasmic reticulum. These observations define TAAR1 as an obligate intracellular target for amphetamines in dopamine neurons and support a model in which distinct pools of TAAR1 mediate the activation of signaling pathways in different compartments to regulate excitatory and dopaminergic neurotransmission.

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  • TAAR1 activation leads to intracellular cAMP signalling that results in PKA and PKC phosphorylation and activation.

    “TAAR1 activation leads to intracellular cAMP signalling that results in PKA and PKC phosphorylation and activation [178–180].”

    From Molecular Characterisation of the Mechanism of Action of Stimulant Drugs Lisdexamfetamine and Methylphenidate on ADHD Neurobiology: A Review (2022) · cites this paper as [179]

    The Introduction reports that TAAR1 activation elevates intracellular cAMP, and the Discussion reports activation of PKA through GαS coupling. The Results describe PKA-mediated phosphorylation of RhoA, not phosphorylation of PKA itself. The cited passages do not support the claim that TAAR1 signalling phosphorylates or activates PKC. No page numbers are provided in our copy. (LLM, conf=0.88)

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Research works printed but not linked: 51

  1. [1]Underhill SM Wheeler DS Li M Watts SD Ingram SL Amara SG Amphetamine modulates excitatory neurotransmission through endocytosis of the glutamate transporter EAAT3 in dopamine neurons Neuron 2014 83 404 16 25033183 PMC4159050
  2. [2]Wheeler DS Underhill SM Stolz DB Murdoch GH Thiels E Romero G Amphetamine activates Rho GTPase signaling to mediate dopamine transporter internalization and acute behavioral effects of amphetamine Proc Natl Acad Sci USA 2015 112 E7138 47 26553986 PMC4697400
  3. [3]Kahlig KM Lute BJ Wei Y Loland CJ Gether U Javitch JA Regulation of dopamine transporter trafficking by intracellular amphetamine Mol Pharmacol 2006 70 542 8 16684900
  4. [4]Borowsky B Adham N Jones KA Raddatz R Artymyshyn R Ogozalek KL Trace amines: identification of a family of mammalian G protein-coupled receptors Proc Natl Acad Sci USA 2001 98 8966 71 11459929 PMC55357
  5. [5]Bunzow JR Sonders MS Arttamangkul S Harrison LM Zhang G Quigley DI Amphetamine, 3,4-methylenedioxymethamphetamine, lysergic acid diethylamide, and metabolites of the catecholamine neurotransmitters are agonists of a rat trace amine receptor Mol Pharm 2001 60 1181 8 11723224
  6. [6]Lindemann L Meyer CA Jeanneau K Bradaia A Ozmen L Bluethmann H Trace amine-associated receptor 1 modulates dopaminergic activity J Pharmacol Exp Ther 2008 324 948 56 18083911
  7. [7]Cotter R Pei Y Mus L Harmeier A Gainetdinov RR Hoener MC The trace amine-associated receptor 1 modulates methamphetamine’s neurochemical and behavioral effects Front Neurosci 2015 9 39 25762894 PMC4327507
  8. [8]Galley G Stalder H Goergler A Hoener MC Norcross RD Optimisation of imidazole compounds as selective TAAR1 agonists: discovery of RO5073012 Bioorg Med Chem Lett 2012 22 5244 8 22795332
  9. [9]Jing L, Zhang Y, Li JX. Effects of the trace amine associated receptor 1 agonist RO5263397 on abuse-related behavioral indices of methamphetamine in rats. Int J Neuropsychopharmacol. 2015;18:1–7. PMC4360231 25522401
  10. [10]Pei Y Asif-Malik A Hoener M Canales JJ A partial trace amine-associated receptor 1 agonist exhibits properties consistent with a methamphetamine substitution treatment Addict Biol 2016 22 1246 56 27193165
  11. [11]Thorn DA Jing L Qiu Y Gancarz-Kausch AM Galuska CM Dietz DM Effects of the trace amine-associated receptor 1 agonist RO5263397 on abuse-related effects of cocaine in rats Neuropsychopharmacology 2014 39 2309 16 24743376 PMC4138753
  12. [12]Thorn DA Zhang C Zhang Y Li JX The trace amine associated receptor 1 agonist RO5263397 attenuates the induction of cocaine behavioral sensitization in rats Neurosci Lett 2014 566 67 71 24561093 PMC3991844
  13. [13]Liu JF, Thorn DA, Zhang Y, Li JX. Effects of trace amine-associated receptor 1 agonists on the expression, reconsolidation, and extinction of cocaine reward memory. Int J Neuropsychopharmacol. 2016:1–9. PMC4966273 26822713
  14. [14]Ran FA Hsu PD Wright J Agarwala V Scott DA Zhang F Genome engineering using the CRISPR-Cas9 system Nat Protoc 2013 8 2281 308 24157548 PMC3969860
  15. [15]Ren XD Kiosses WB Schwartz MA Regulation of the small GTP-binding protein Rho by cell adhesion and the cytoskeleton EMBO J 1999 18 578 85 9927417 PMC1171150
  16. [16]Depry C Allen MD Zhang J Visualization of PKA activity in plasma membrane microdomains Mol Biosyst 2011 7 52 8 20838685
  17. [17]Herbst KJ Allen MD Zhang J Spatiotemporally regulated protein kinase A activity is a critical regulator of growth factor-stimulated extracellular signal-regulated kinase signaling in PC12 cells Mol Cell Biol 2011 31 4063 75 21807900 PMC3187359
  18. [18]Mehta S Aye-Han NN Ganesan A Oldach L Gorshkov K Zhang J Calmodulin-controlled spatial decoding of oscillatory Ca2+ signals by calcineurin Elife 2014 3 e03765 25056880 PMC4141273
  19. [19]Gilchrist A Li A Hamm HE Design and use of C-terminal minigene vectors for studying role of heterotrimeric G proteins Methods Enzymol 2002 344 58 69 11771412
  20. [20]Zimmermann L Stephens A Nam SZ Rau D Kubler J Lozajic M A completely reimplemented MPI bioinformatics toolkit with a new HHpred server at its core J Mol Biol 2018 430 2237 43 29258817
  21. [21]Pertz O Hodgson L Klemke RL Hahn KM Spatiotemporal dynamics of RhoA activity in migrating cells Nature 2006 440 1069 72 16547516
  22. [22]Rizzo MA Springer GH Granada B Piston DW An improved cyan fluorescent protein variant useful for FRET Nat Biotechnol 2004 22 445 9 14990965
  23. [23]Porzgen P Park SK Hirsh J Sonders MS Amara SG The antidepressant-sensitive dopamine transporter in Drosophila melanogaster: a primordial carrier for catecholamines Mol Pharmacol 2001 59 83 95 11125028
  24. [24]Alouf JE Geoffroy C Production, purification, and assay of streptolysin O Methods Enzymol 1988 165 52 9 3231118
  25. [25]Revel FG Moreau JL Gainetdinov RR Bradaia A Sotnikova TD Mory R TAAR1 activation modulates monoaminergic neurotransmission, preventing hyperdopaminergic, and hypoglutamatergic activity Proc Natl Acad Sci USA 2011 108 8485 90 21525407 PMC3101002
  26. [26]Revel FG Moreau JL Gainetdinov RR Ferragud A Velazquez-Sanchez C Sotnikova TD Trace amine-associated receptor 1 partial agonism reveals novel paradigm for neuropsychiatric therapeutics Biol Psychiatry 2012 72 934 42 22705041
  27. [27]Xie Z Miller GM A receptor mechanism for methamphetamine action in dopamine transporter regulation in brain J Pharmacol Exp Ther 2009 330 316 25 19364908 PMC2700171
  28. [28]Ellerbroek SM Wennerberg K Burridge K Serine phosphorylation negatively regulates RhoA in vivo J Biol Chem 2003 278 19023 31 12654918
  29. [29]Ni Q Titov DV Zhang J Analyzing protein kinase dynamics in living cells with FRET reporters Methods 2006 40 279 86 16908183
  30. [30]Ponsioen B Zhao J Riedl J Zwartkruis F van der Krogt G Zaccolo M Detecting cAMP-induced Epac activation by fluorescence resonance energy transfer: Epac as a novel cAMP indicator Embo Rep 2004 5 1176 80 15550931 PMC1299185
  31. [31]Siehler S Regulation of RhoGEF proteins by G12/13-coupled receptors Br J Pharmacol 2009 158 41 9 19226283 PMC2795247
  32. [32]Momotani K Somlyo AV p63RhoGEF: a new switch for G(q)-mediated activation of smooth muscle Trends Cardiovasc Med 2012 22 122 7 22902181 PMC3472095
  33. [33]Devost D Sleno R Petrin D Zhang A Shinjo Y Okde R Conformational profiling of the AT1 angiotensin II receptor reflects biased agonism, G protein coupling, and cellular context J Biol Chem 2017 292 5443 56 28213525 PMC5392687
  34. [34]Hu J Stern M Gimenez LE Wanka L Zhu L Rossi M A G protein-biased designer G protein-coupled receptor useful for studying the physiological relevance of Gq/11-dependent signaling pathways J Biol Chem 2016 291 7809 20 26851281 PMC4824988
  35. [35]Stallaert W van der Westhuizen ET Schonegge AM Plouffe B Hogue M Lukashova V Purinergic receptor transactivation by the beta2-adrenergic receptor increases intracellular Ca2+ in nonexcitable cells Mol Pharmacol 2017 91 533 44 28280061
  36. [36]Gilchrist A Bunemann M Li A Hosey MM Hamm HE A dominant-negative strategy for studying roles of G proteins in vivo J Biol Chem 1999 274 6610 6 10037756
  37. [37]Gilchrist A Vanhauwe JF Li A Thomas TO Voyno-Yasenetskaya T Hamm HE G alpha minigenes expressing C-terminal peptides serve as specific inhibitors of thrombin-mediated endothelial activation J Biol Chem 2001 276 25672 9 11274183
  38. [38]Rasmussen SG DeVree BT Zou Y Kruse AC Chung KY Kobilka TS Crystal structure of the beta2 adrenergic receptor-Gs protein complex Nature 2011 477 549 55 21772288 PMC3184188
  39. [39]Li MH Underhill SM Reed C Phillips TJ Amara SG Ingram SL Amphetamine and methamphetamine increase NMDAR-GluN2B synaptic currents in midbrain dopamine neurons Neuropsychopharmacology 2017 42 1539 47 27976681 PMC5436114
  40. [40]Calebiro D Nikolaev VO Persani L Lohse MJ Signaling by internalized G-protein-coupled receptors Trends Pharmacol Sci 2010 31 221 8 20303186
  41. [42]Katritch V Cherezov V Stevens RC Structure-function of the G protein-coupled receptor superfamily Annu Rev Pharmacol Toxicol 2013 53 531 56 23140243 PMC3540149
  42. [43]Berry MD Shitut MR Almousa A Alcorn J Tomberli B Membrane permeability of trace amines: evidence for a regulated, activity-dependent, nonexocytotic, synaptic release Synapse 2013 67 656 67 23564683
  43. [44]Gulaboski R Cordeiro MN Milhazes N Garrido J Borges F Jorge M Evaluation of the lipophilic properties of opioids, amphetamine-like drugs, and metabolites through electrochemical studies at the interface between two immiscible solutions Anal Biochem 2007 361 236 43 17178092
  44. [45]Tanabe H Fujii Y Okada-Iwabu M Iwabu M Nakamura Y Hosaka T Crystal structures of the human adiponectin receptors Nature 2015 520 312 6 25855295 PMC4477036
  45. [46]Purgert CA Izumi Y Jong YJ Kumar V Zorumski CF O’Malley KL Intracellular mGluR5 can mediate synaptic plasticity in the hippocampus J Neurosci 2014 34 4589 98 24672004 PMC3965784
  46. [47]Gasser PJ Hurley MM Chan J Pickel VM Organic cation transporter 3 (OCT3) is localized to intracellular and surface membranes in select glial and neuronal cells within the basolateral amygdaloid complex of both rats and mice Brain Struct Funct 2017 222 1913 28 27659446 PMC5362368
  47. [48]Berry MD Hart S Pryor AR Hunter S Gardiner D Pharmacological characterization of a high-affinity p-tyramine transporter in rat brain synaptosomes Sci Rep 2016 6 38006 27901065 PMC5128819
  48. [49]Yen HY Hoi KK Liko I Hedger G Horrell MR Song W PtdIns(4,5)P2 stabilizes active states of GPCRs and enhances selectivity of G-protein coupling Nature 2018 559 423 7 29995853 PMC6059376
  49. [50]Oldham WM Hamm HE Heterotrimeric G protein activation by G-protein-coupled receptors Nat Rev Mol Cell Biol 2008 9 60 71 18043707
  50. [51]Lohani S Martig AK Underhill SM DeFrancesco A Roberts MJ Rinaman L Burst activation of dopamine neurons produces prolonged post-burst availability of actively released dopamine Neuropsychopharmacology 2018 43 2083 92 29795245 PMC6098082
  51. [52]Zigmond MJ Hastings TG Perez RG Increased dopamine turnover after partial loss of dopaminergic neurons: compensation or toxicity? Park Relat Disord 2002 8 389 93 12217625

Cited by

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Authors
Underhill, S. M., Hullihen, P. D., Chen, J., Fenollar-Ferrer, C., Rizzo, M. A., Ingram, S. L., & Amara, S. G.
Affiliations
  • National Institutes of Health (US)
  • University of Maryland, Baltimore (US)
  • Oregon Health & Science University (US)
Corresponding author
Underhill, Suzanne M.
ORCID
Venue
Molecular Psychiatry
Publisher
Springer Science and Business Media LLC
Published
2019-08-09
Volume
26
Issue
4
Pages
1208-1223
ISSN
  • 1359-4184 (ISSN-L)
  • 1476-5578
DOI
10.1038/s41380-019-0469-2
PMC
PMC7038576
OpenAlex
W2967836394
Cited by
92
References
1
Citations by year
2026: 12 · 2025: 13 · 2024: 9 · 2023: 14 · 2022: 21 · 2021: 11 · 2020: 9 · 2019: 3
FWCI
5.25
Type
journal article
Language
en
Open access
Licence
Locations
  • Molecular Psychiatry (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
NIMH Intramural Research
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neurotransmitter Receptor Influence on Behavior
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
Keywords
amphetamine, TAAR1, Gα13, Gs protein, RhoA signaling, protein kinase A, dopaminergic neurons, endoplasmic reticulum, attention deficit hyperactivity disorder, FRET biosensor, G protein signaling
MeSH terms
Trace Amine-Associated Receptors, Amphetamine, Animals, Chromogranins, Dopamine, Synaptic Transmission, GTP-Binding Protein alpha Subunits, Gs, Receptors, G-Protein-Coupled, GTP-Binding Protein alpha Subunits, G12-G13, Mice, Dopaminergic Neurons

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