Inhibition of G-Protein-Activated Inwardly Rectifying K+ Channels by the Selective Norepinephrine Reuptake Inhibitors Atomoxetine and Reboxetine

Kobayashi, Toru; Washiyama, Kazuo; Ikeda, Kazutaka

doi:10.1038/npp.2010.27

Original Article
Published: 10 March 2010

Inhibition of G-Protein-Activated Inwardly Rectifying K⁺ Channels by the Selective Norepinephrine Reuptake Inhibitors Atomoxetine and Reboxetine

Toru Kobayashi¹,
Kazuo Washiyama¹ &
Kazutaka Ikeda²

Neuropsychopharmacology volume 35, pages 1560–1569 (2010)Cite this article

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Abstract

Atomoxetine and reboxetine are commonly used as selective norepinephrine reuptake inhibitors (NRIs) for the treatment of attention-deficit/hyperactivity disorder and depression, respectively. Furthermore, recent studies have suggested that NRIs may be useful for the treatment of several other psychiatric disorders. However, the molecular mechanisms underlying the various effects of NRIs have not yet been sufficiently clarified. G-protein-activated inwardly rectifying K⁺ (GIRK or Kir3) channels have an important function in regulating neuronal excitability and heart rate, and GIRK channel modulation has been suggested to be a potential treatment for several neuropsychiatric disorders and cardiac arrhythmias. In this study, we investigated the effects of atomoxetine and reboxetine on GIRK channels using the Xenopus oocyte expression assay. In oocytes injected with mRNA for GIRK1/GIRK2, GIRK2, or GIRK1/GIRK4 subunits, extracellular application of atomoxetine or reboxetine reversibly reduced GIRK currents. The inhibitory effects were concentration-dependent, but voltage-independent, and time-independent during each voltage pulse. However, Kir1.1 and Kir2.1 channels were insensitive to atomoxetine and reboxetine. Atomoxetine and reboxetine also inhibited GIRK currents induced by activation of cloned A₁ adenosine receptors or by intracellularly applied GTPγS, a nonhydrolyzable GTP analogue. Furthermore, the GIRK currents induced by ethanol were concentration-dependently inhibited by extracellularly applied atomoxetine but not by intracellularly applied atomoxetine. The present results suggest that atomoxetine and reboxetine inhibit brain- and cardiac-type GIRK channels, revealing a novel characteristic of clinically used NRIs. GIRK channel inhibition may contribute to some of the therapeutic effects of NRIs and adverse side effects related to nervous system and heart function.

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References

Ahern TH, Javors MA, Eagles DA, Martillotti J, Mitchell HA, Liles LC et al (2006). The effects of chronic norepinephrine transporter inactivation on seizure susceptibility in mice. Neuropsychopharmacology 31: 730–738.
Article CAS Google Scholar
Barker MJ, Benitez JG, Ternullo S, Juhl GA (2004). Acute oxcarbazepine and atomoxetine overdose with quetiapine. Vet Hum Toxicol 46: 130–132.
PubMed Google Scholar
Belle DJ, Ernest CS, Sauer JM, Smith BP, Thomasson HR, Witcher JW (2002). Effect of potent CYP2D6 inhibition by paroxetine on atomoxetine pharmacokinetics. J Clin Pharmacol 42: 1219–1227.
Article CAS Google Scholar
Bettahi I, Marker CL, Roman MI, Wickman K (2002). Contribution of the Kir3.1 subunit to the muscarinic-gated atrial potassium channel IKACh. J Biol Chem 277: 48282–48288.
Article CAS Google Scholar
Blednov YA, Stoffel M, Chang SR, Harris RA (2001). Potassium channels as targets for ethanol: studies of G-protein-coupled inwardly rectifying potassium channel 2 (GIRK2) null mutant mice. J Pharmacol Exp Ther 298: 521–530.
CAS PubMed Google Scholar
Boot J, Cases M, Clark BP, Findlay J, Gallagher PT, Hayhurst L et al (2005). Discovery and structure-activity relationships of novel selective norepinephrine and dual serotonin/norepinephrine reuptake inhibitors. Bioorg Med Chem Lett 15: 699–703.
Article CAS Google Scholar
Cruz HG, Berton F, Sollini M, Blanchet C, Pravetoni M, Wickman K et al (2008). Absence and rescue of morphine withdrawal in KIR/Kir3 knock-out mice. J Neurosci 28: 4069–4077.
Article CAS Google Scholar
Cusack B, Nelson A, Richelson E (1994). Binding of antidepressants to human brain receptors: focus on newer generation compounds. Psychopharmacology 114: 559–565.
Article CAS Google Scholar
Dascal N (1997). Signalling via the G protein-activated K⁺ channels. Cell Signal 9: 551–573.
Article CAS Google Scholar
Fraser SP, Djamgoz MBA (1992). Xenopus oocytes: endogenous electrophysiological characteristics. In: Osborne NN (ed). Current Aspects of the Neurosciences, Vol 4. Macmillan: London. pp 267–315.
Chapter Google Scholar
Garland M, Kirkpatrick P (2004). Atomoxetine hydrochloride. Nat Rev Drug Discovery 3: 385–386.
Article CAS Google Scholar
Garside D, Ropero-Miller JD, Riemer EC (2006). Postmortem tissue distribution of atomoxetine following fatal and nonfatal doses: three case reports. J Forensic Sci 51: 179–182.
Article CAS Google Scholar
Geller D, Donnelly C, Lopez F, Rubin R, Newcorn J, Sutton V et al (2007). Atomoxetine treatment for pediatric patients with attention-deficit/hyperactivity disorder with comorbid anxiety disorder. J Am Acad Child Adolesc Psychiatry 46: 1119–1127.
Article Google Scholar
Hajós M, Fleishaker JC, Filipiak-Reisner JK, Brown MT, Wong EHF (2004). The selective norepinephrine reuptake inhibitor antidepressant reboxetine: pharmacological and clinical profile. CNS Drug Rev 10: 23–44.
Article Google Scholar
Hashimoto N, Yamashita T, Tsuruzoe N (2006). Tertiapine, a selective IKACh blocker, terminates atrial fibrillation with selective atrial effective refractory period prolongation. Pharmacol Res 54: 136–141.
Article CAS Google Scholar
Hill KG, Alva H, Blednov YA, Cunningham CL (2003). Reduced ethanol-induced conditioned taste aversion and conditioned place preference in GIRK2 null mutant mice. Psychopharmacology 169: 108–114.
Article CAS Google Scholar
Ho K, Nichols CG, Lederer WJ, Lytton J, Vassilev PM, Kanazirska MV et al (1993). Cloning and expression of an inwardly rectifying ATP-regulated potassium channel. Nature 362: 31–38.
Article CAS Google Scholar
Ikeda K, Yoshii M, Sora I, Kobayashi T (2003). Opioid receptor coupling to GIRK channels: in vitro studies using a Xenopus oocyte expression system and in vivo studies on weaver mutant mice. Methods Mol Med 84: 53–64.
CAS PubMed Google Scholar
Inanobe A, Yoshimoto Y, Horio Y, Morishige K-I, Hibino H, Matsumoto S et al (1999). Characterization of G-protein-gated K⁺ channels composed of Kir3.2 subunits in dopaminergic neurons of the substantia nigra. J Neurosci 19: 1006–1017.
Article CAS Google Scholar
Kadhe NG, Chillar AJ, Deshmukh YA (2003). Reboxetine: a novel antidepressant. J Postgrad Med 49: 373–375.
CAS PubMed Google Scholar
Kanegawa N, Kiyono Y, Kimura H, Sugita T, Kajiyama S, Kawashima H et al (2006). Synthesis and evaluation of radioiodinated (S,S)-2-(α-(2-iodophenoxy)benzyl)morpholine for imaging brain norepinephrine transporter. Eur J Nucl Med Mol Imaging 33: 639–647.
Article CAS Google Scholar
Karschin C, Dißmann E, Stuhmer W, Karschin A (1996). IRK(1-3) and GIRK(1-4) inwardly rectifying K⁺ channel mRNAs are differentially expressed in the adult rat brain. J Neurosci 16: 3559–3570.
Article CAS Google Scholar
Keller NR, Diedrich A, Appalsamy M, Tuntrkool S, Lonce S, Finney C et al (2004). Norepinephrine transporter-deficient mice exhibit excessive tachycardia and elevated blood pressure with wakefulness and activity. Circulation 110: 1191–1196.
Article CAS Google Scholar
Kiyono Y, Kanegawa N, Kawashima H, Kitamura Y, Iida Y, Saji H (2004). Evaluation of radioiodinated (R)-N-methyl-3-(2-iodophenoxy)-3-phenylpropanamine as a ligand for brain norepinephrine transporter imaging. Nucl Med Biol 31: 147–153.
Article CAS Google Scholar
Kiyono Y, Sugita T, Ueda M, Kawashima H, Kanegawa N, Kuge Y et al (2008). Evaluation of radioiodinated (2S,αS)-2-(α-(2-iodophenoxy)benzyl)morpholine as a radioligand for imaging of norepinephrine transporter in the heart. Nucl Med Biol 35: 213–218.
Article CAS Google Scholar
Kobayashi T, Ikeda K (2006). G protein-activated inwardly rectifying potassium channels as potential therapeutic targets. Curr Pharm Des 12: 4513–4523.
Article CAS Google Scholar
Kobayashi T, Ikeda K, Ichikawa T, Abe S, Togashi S, Kumanishi T (1995). Molecular cloning of a mouse G-protein-activated K⁺ channel (mGIRK1) and distinct distributions of three GIRK (GIRK1, 2 and 3) mRNAs in mouse brain. Biochem Biophys Res Commun 208: 1166–1173.
Article CAS Google Scholar
Kobayashi T, Ikeda K, Kojima H, Niki H, Yano R, Yoshioka T et al (1999). Ethanol opens G-protein-activated inwardly rectifying K⁺ channels. Nat Neurosci 2: 1091–1097.
Article CAS Google Scholar
Kobayashi T, Ikeda K, Kumanishi T (2000). Inhibition by various antipsychotic drugs of the G-protein-activated inwardly rectifying K⁺ (GIRK) channels expressed in Xenopus oocytes. Br J Pharmacol 129: 1716–1722.
Article CAS Google Scholar
Kobayashi T, Ikeda K, Kumanishi T (2002). Functional characterization of an endogenous Xenopus oocyte adenosine receptor. Br J Pharmacol 135: 313–322.
Article CAS Google Scholar
Kobayashi T, Washiyama K, Ikeda K (2003). Inhibition of G protein-activated inwardly rectifying K⁺ channels by fluoxetine (Prozac). Br J Pharmacol 138: 1119–1128.
Article CAS Google Scholar
Kobayashi T, Washiyama K, Ikeda K (2006). Inhibition of G protein-activated inwardly rectifying K⁺ channels by the antidepressant paroxetine. J Pharmacol Sci 102: 278–287.
Article CAS Google Scholar
Kovoor A, Henry DJ, Chavkin C (1995). Agonist-induced desensitization of the mu opioid receptor-coupled potassium channel (GIRK1). J Biol Chem 270: 589–595.
Article CAS Google Scholar
Kovoor P, Wickman K, Maguire CT, Pu W, Gehrmann J, Berul CI et al (2001). Evaluation of the role of IKACh in atrial fibrillation using a mouse knockout model. J Am Coll Cardiol 37: 2136–2143.
Article CAS Google Scholar
Krapivinsky G, Gordon EA, Wickman K, Velimirovic B, Krapivinsky L, Clapham DE (1995). The G-protein-gated atrial K⁺ channel IKACh is a heteromultimer of two inwardly rectifying K⁺-channel proteins. Nature 374: 135–141.
Article CAS Google Scholar
Kubo Y, Baldwin TJ, Jan YN, Jan LY (1993a). Primary structure and functional expression of a mouse inward rectifier potassium channel. Nature 362: 127–133.
Article CAS Google Scholar
Kubo Y, Reuveny E, Slesinger PA, Jan YN, Jan LY (1993b). Primary structure and functional expression of a rat G-protein-coupled muscarinic potassium channel. Nature 364: 802–806.
Article CAS Google Scholar
Kuzhikandathil EV, Oxford GS (2002). Classic D1 dopamine receptor antagonist R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine hydrochloride (SCH23390) directly inhibits G protein-coupled inwardly rectifying potassium channels. Mol Pharmacol 62: 119–126.
Article CAS Google Scholar
Lesage F, Guillemare E, Fink M, Duprat F, Heurteaux C, Fosset M et al (1995). Molecular properties of neuronal G-protein-activated inwardly rectifying K⁺ channels. J Biol Chem 270: 28660–28667.
Article CAS Google Scholar
Lewohl JM, Wilson WR, Mayfield RD, Brozowski SJ, Morrisett RA, Harris RA (1999). G-protein-coupled inwardly rectifying potassium channels are targets of alcohol action. Nat Neurosci 2: 1084–1090.
Article CAS Google Scholar
Liao YJ, Jan YN, Jan LY (1996). Heteromultimerization of G-protein-gated inwardly rectifying K⁺ channel proteins GIRK1 and GIRK2 and their altered expression in weaver brain. J Neurosci 16: 7137–7150.
Article CAS Google Scholar
LoVecchio F, Kashani J (2006). Isolated atomoxetine (STRATTERA^TM) ingestions commonly result in toxicity. J Emerg Med 31: 267–268.
Article Google Scholar
Lüscher C, Jan LY, Stoffel M, Malenka RC, Nicoll RA (1997). G protein-coupled inwardly rectifying K⁺ channels (GIRKs) mediate postsynaptic but not presynaptic transmitter actions in hippocampal neurons. Neuron 19: 687–695.
Article Google Scholar
McElroy SL, Guerdjikova A, Kotwal R, Welge JA, Nelson EB, Lake KA et al (2007). Atomoxetine in the treatment of binge-eating disorder: a randomized placebo-controlled trial. J Clin Psychiatry 68: 390–398.
Article CAS Google Scholar
Montgomery SA (2005). Antidepressants and seizures: emphasis on newer agents and clinical implications. Int J Clin Pract 59: 1435–1440.
Article CAS Google Scholar
Morgan AD, Carroll ME, Loth AK, Stoffel M, Wickman K (2003). Decreased cocaine self-administration in Kir3 potassium channel subunit knockout mice. Neuropsychopharmacology 28: 932–938.
Article CAS Google Scholar
North RA (1989). Drug receptors and the inhibition of nerve cells. Br J Pharmacol 98: 13–28.
Article CAS Google Scholar
Öhman D, Norlander B, Peterson C, Bengtsson F (2001). Bioanalysis of racemic reboxetine and its desethylated metabolite in a therapeutic drug monitoring setting using solid phase extraction and HPLC. Ther Drug Monit 23: 27–34.
Article Google Scholar
Poggesi I, Pellizzoni C, Fleishaker JC (2000). Pharmacokinetics of reboxetine in elderly patients with depressive disorders. Int J Clin Pharmacol Ther 38: 254–259.
Article CAS Google Scholar
Reimann F, Ashcroft FM (1999). Inwardly rectifying potassium channels. Curr Opin Cell Biol 11: 503–508.
Article CAS Google Scholar
Sawant S, Daviss SR (2004). Seizures and prolonged QTc with atomoxetine overdose. Am J Psychiatry 161: 757.
Article Google Scholar
Scherer D, Hassel D, Bloehs R, Zitron E, von Löwenstern K, Seyler C et al (2009). Selective noradrenaline reuptake inhibitor atomoxetine directly blocks hERG currents. Br J Pharmacol 156: 226–236.
Article CAS Google Scholar
Signorini S, Liao YJ, Duncan SA, Jan LY, Stoffel M (1997). Normal cerebellar development but susceptibility to seizures in mice lacking G protein-coupled, inwardly rectifying K⁺ channel GIRK2. Proc Natl Acad Sci USA 94: 923–927.
Article CAS Google Scholar
Simpson D, Plosker GL (2004). Atomoxetine a review of its use in adults with attention deficit hyperactivity disorder. Drugs 64: 205–222.
Article CAS Google Scholar
Szerman N, Peris L, Mesías B, Colis P, Rosa J, Prieto A et al (2005). Reboxetine for the treatment of patients with cocaine dependence disorder. Hum Psychopharmacol Clin Exp 20: 189–192.
Article CAS Google Scholar
Takahashi T, Kobayashi T, Ozaki M, Takamatsu Y, Ogai Y, Ohta M et al (2006). G protein-activated inwardly rectifying K⁺ channel inhibition and rescue of weaver mouse motor functions by antidepressants. Neurosci Res 54: 104–111.
Article CAS Google Scholar
Tirado CF, Goldman M, Lynch K, Kampman KM, Obrien CP (2008). Atomoxetine for treatment of marijuana dependence: a report on the efficacy and high incidence of gastrointestinal adverse events in a pilot study. Drug Alcohol Depend 94: 254–257.
Article CAS Google Scholar
Versiani M, Cassano G, Perugi G, Benedetti A, Mastalli L, Nardi A et al (2002). Reboxetine, a selective norepinephrine reuptake inhibitor, is an effective and well-tolerated treatment for panic disorder. J Clin Psychiatry 63: 31–37.
Article CAS Google Scholar
Weber WM (1999). Ion currents of Xenopus laevis oocytes: state of the art. Biochim Biophys Acta 1421: 213–233.
Article CAS Google Scholar
Wernicke JF, Holdridge KC, Jin L, Edison T, Zhang S, Bangs ME et al (2007). Seizure risk in patients with attention-deficit-hyperactivity disorder treated with atomoxetine. Dev Med Child Neurol 49: 498–502.
Article Google Scholar
Wilens TE, Adler LA, Weiss MD, Michelson D, Ramsey JL, Moore RJ et al (2008). Atomoxetine treatment of adults with ADHD and comorbid alcohol use disorders. Drug Alcohol Depend 96: 145–154.
Article CAS Google Scholar
Witcher JW, Long A, Smith B, Sauer JM, Heilgenstein J, Wilens T et al (2003). Atomoxetine pharmacokinetics in children and adolescents with attention deficit hyperactivity disorder. J Child Adolesc Psychopharmacol 13: 53–64.
Article Google Scholar
Wong EHF, Sonders MS, Amara SG, Tinholt PM, Piercey MFP, Hoffmann WP et al (2000). Reboxetine: a pharmacologically potent, selective, and specific norepinephrine reuptake inhibitor. Biol Psychiatry 47: 818–829.
Article CAS Google Scholar

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Acknowledgements

We are grateful to Dr Kansaku Baba for his cooperation and Mr Kazuo Kobayashi (Niigata University) for his assistance. We also thank Dr Steven C Hebert and Dr Lily Y Jan for generously providing the Kir1.1 cDNA and Kir2.1 cDNA, respectively. This work was supported by research grants from the Ministry of Education, Science, Sports, and Culture of Japan and from the Ministry of Health, Labour, and Welfare of Japan.

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Department of Molecular Neuropathology, Brain Research Institute, Niigata University, Chuo-ku, Niigata, Niigata, Japan
Toru Kobayashi & Kazuo Washiyama
Division of Psychobiology, Tokyo Institute of Psychiatry, Setagaya-ku, Tokyo, Japan
Kazutaka Ikeda

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Toru Kobayashi
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Kazuo Washiyama
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Kazutaka Ikeda
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Correspondence to Toru Kobayashi.

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The authors declare that over the past 3 years Kazutaka Ikeda has received research grants or expenses that are not related to this study from Fujifilm Corporation, the Mitsubishi Foundation, the Naito Foundation, and the Smoking Science Foundation, and a lecture fee from Dainippon Sumitomo Pharma and Kyowa Hakko Kirin. The authors declare that, except for income received from their primary employer and the aforementioned disclosures, no financial support or compensation has been received from any individual or corporate entity over the past 3 years for research or professional service, and there are no personal financial holdings that could be perceived as constituting a potential conflict of interest.

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Kobayashi, T., Washiyama, K. & Ikeda, K. Inhibition of G-Protein-Activated Inwardly Rectifying K⁺ Channels by the Selective Norepinephrine Reuptake Inhibitors Atomoxetine and Reboxetine. Neuropsychopharmacol 35, 1560–1569 (2010). https://doi.org/10.1038/npp.2010.27

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Received: 04 November 2009
Revised: 30 January 2010
Accepted: 08 February 2010
Published: 10 March 2010
Issue date: June 2010
DOI: https://doi.org/10.1038/npp.2010.27

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