title: Mechanisms of amphetamine action illuminated through optical monitoring of dopamine synaptic vesicles in Drosophila brain.
abstract: Amphetamines elevate extracellular dopamine, but the underlying mechanisms remain uncertain. Here we show in rodents that acute pharmacological inhibition of the vesicular monoamine transporter (VMAT) blocks amphetamine-induced locomotion and self-administration without impacting cocaine-induced behaviours. To study VMAT's role in mediating amphetamine action in dopamine neurons, we have used novel genetic, pharmacological and optical approaches in Drosophila melanogaster. In an ex vivo whole-brain preparation, fluorescent reporters of vesicular cargo and of vesicular pH reveal that amphetamine redistributes vesicle contents and diminishes the vesicle pH-gradient responsible for dopamine uptake and retention. This amphetamine-induced deacidification requires VMAT function and results from net H(+) antiport by VMAT out of the vesicle lumen coupled to inward amphetamine transport. Amphetamine-induced vesicle deacidification also requires functional dopamine transporter (DAT) at the plasma membrane. Thus, we find that at pharmacologically relevant concentrations, amphetamines must be actively transported by DAT and VMAT in tandem to produce psychostimulant effects.
authors.affiliation: Department of Psychiatry, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA.
authors.forename: Zachary
authors.initials: Z
authors.lastname: Freyberg
authors.affiliation: Department of Psychiatry, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA.|Department of Neurology, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Sonders
authors.affiliation: Department of Psychiatry, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA.
authors.forename: Jenny I
authors.initials: JI
authors.lastname: Aguilar
authors.affiliation: Psychobiology Section, Intramural Research Program, Department of Health and Human Services, National Institute on Drug Abuse, National Institutes of Health, Baltimore, Maryland 21224, USA.
authors.forename: Takato
authors.initials: T
authors.lastname: Hiranita
authors.affiliation: Department of Psychiatry, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA.
authors.forename: Caline S
authors.initials: CS
authors.lastname: Karam
authors.affiliation: Department of Biochemistry and Molecular Biophysics, Howard Hughes Medical Institute, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.
authors.forename: Jorge
authors.initials: J
authors.lastname: Flores
authors.affiliation: Department of Psychiatry, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA.
authors.forename: Andrea B
authors.initials: AB
authors.lastname: Pizzo
authors.affiliation: Department of Psychiatry, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA.
authors.forename: Yuchao
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Psychiatry, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA.
authors.forename: Zachary J
authors.initials: ZJ
authors.lastname: Farino
authors.affiliation: Department of Psychiatry and Biobehavioral Sciences and Semel Institute for Neuroscience and Human Behavior, Hatos Center for Neuropharmacology, David Geffen School of Medicine University of California, Los Angeles, California 90095, USA.
authors.forename: Audrey
authors.initials: A
authors.lastname: Chen
authors.affiliation: UCLA Interdepartmental Program in Molecular Toxicology, University of California, Los Angeles, California 90095, USA.
authors.forename: Ciara A
authors.initials: CA
authors.lastname: Martin
authors.affiliation: Psychobiology Section, Intramural Research Program, Department of Health and Human Services, National Institute on Drug Abuse, National Institutes of Health, Baltimore, Maryland 21224, USA.
authors.forename: Theresa A
authors.initials: TA
authors.lastname: Kopajtic
authors.affiliation: Department of Psychiatry and Biobehavioral Sciences and Semel Institute for Neuroscience and Human Behavior, Hatos Center for Neuropharmacology, David Geffen School of Medicine University of California, Los Angeles, California 90095, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Fei
authors.affiliation: Department of Chemistry, Columbia University, New York, New York 10027, USA.
authors.forename: Gang
authors.initials: G
authors.lastname: Hu
authors.affiliation: School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Republic of China 10055.
authors.forename: Yi-Ying
authors.initials: YY
authors.lastname: Lin
authors.affiliation: Department of Neurology, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.
authors.forename: Eugene V
authors.initials: EV
authors.lastname: Mosharov
authors.affiliation: Center for Motor Neuron Biology and Disease, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Department of Pathology and Cell Biology, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Department of Neuroscience, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.
authors.forename: Brian D
authors.initials: BD
authors.lastname: McCabe
authors.affiliation: Department of Psychology, Stern College for Women, Yeshiva University, New York, New York 10016, USA.
authors.forename: Robin
authors.initials: R
authors.lastname: Freyberg
authors.affiliation: Department of Chemistry, Wichita State University, Wichita, Kansas 67260, USA.
authors.forename: Kandatege
authors.initials: K
authors.lastname: Wimalasena
authors.affiliation: School of Pharmacy, College of Medicine, National Taiwan University, Taipei, Republic of China 10055.
authors.forename: Ling-Wei
authors.initials: LW
authors.lastname: Hsin
authors.affiliation: Department of Chemistry, Columbia University, New York, New York 10027, USA.
authors.forename: Dalibor
authors.initials: D
authors.lastname: Sames
authors.affiliation: Department of Psychiatry and Biobehavioral Sciences and Semel Institute for Neuroscience and Human Behavior, Hatos Center for Neuropharmacology, David Geffen School of Medicine University of California, Los Angeles, California 90095, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Krantz
authors.affiliation: Psychobiology Section, Intramural Research Program, Department of Health and Human Services, National Institute on Drug Abuse, National Institutes of Health, Baltimore, Maryland 21224, USA.
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Katz
authors.affiliation: Department of Psychiatry, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA.|Department of Neurology, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Department of Pharmacology, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.
authors.forename: David
authors.initials: D
authors.lastname: Sulzer
authors.affiliation: Department of Psychiatry, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.|Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, New York 10032, USA.|Department of Pharmacology, College of Physicians &Surgeons, Columbia University, New York, New York 10032, USA.
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Javitch
mesh_terms: D000661:Amphetamine / Q000494:pharmacology*; D000818:Animals; D030801:Animals, Genetically Modified; D001921:Brain / Q000187:drug effects* / Q000378:metabolism; D003042:Cocaine / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D015259:Dopamine Agents / Q000494:pharmacology*; D050483:Dopamine Plasma Membrane Transport Proteins / Q000187:drug effects* / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000187:drug effects* / Q000378:metabolism; D004331:Drosophila melanogaster; D057809:HEK293 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008124:Locomotion / Q000187:drug effects*; D008694:Methamphetamine / Q000494:pharmacology; D008774:Methylphenidate / Q000494:pharmacology; D061848:Optical Imaging; D051381:Rats; D013572:Synaptic Vesicles / Q000187:drug effects*; D050493:Vesicular Monoamine Transport Proteins / Q000037:antagonists & inhibitors* / Q000187:drug effects / Q000378:metabolism
chemical_list: D015259:Dopamine Agents; D050483:Dopamine Plasma Membrane Transport Proteins; C493454:Slc18a2 protein, rat; D050493:Vesicular Monoamine Transport Proteins; D008774:Methylphenidate; D008694:Methamphetamine; D000661:Amphetamine; D003042:Cocaine; D004298:Dopamine