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Amphetamine and methamphetamine differentially affect dopamine transporters in vitro and in vivo.
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The psychostimulants d-amphetamine (AMPH) and methamphetamine (METH) release excess dopamine (DA) into the synaptic clefts of dopaminergic neurons. Abnormal DA release is thought to occur by reverse transport through the DA transporter (DAT), and it is believed to underlie the severe behavioral effects of these drugs. Here we compare structurally similar AMPH and METH on DAT function in a heterologous expression system and in an animal model. In the in vitro expression system, DAT-mediated whole-cell currents were greater for METH stimulation than for AMPH. At the same voltage and concentration, METH released five times more DA than AMPH and did so at physiological membrane potentials. At maximally effective concentrations, METH released twice as much [Ca(2+)](i) from internal stores compared with AMPH. [Ca(2+)](i) responses to both drugs were independent of membrane voltage but inhibited by DAT antagonists. Intact phosphorylation sites in the N-terminal domain of DAT were required for the AMPH- and METH-induced increase in [Ca(2+)](i) and for the enhanced effects of METH on [Ca(2+)](i) elevation. Calmodulin-dependent protein kinase II and protein kinase C inhibitors alone or in combination also blocked AMPH- or METH-induced Ca(2+) responses. Finally, in the rat nucleus accumbens, in vivo voltammetry showed that systemic application of METH inhibited DAT-mediated DA clearance more efficiently than AMPH, resulting in excess external DA. Together these data demonstrate that METH has a stronger effect on DAT-mediated cell physiology than AMPH, which may contribute to the euphoric and addictive properties of METH compared with AMPH.
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“D-amphetamine and methamphetamine enter brain rapidly after administration. They competitively inhibit dopamine transporter (DAT) clearing dopamine (DA), and also release DA via reversing DAT direction, ultimately causing excessive DA accumulation in synaptic cleft ultimately (11).”
From Effects of Lisdexamfetamine, a Prodrug of D-Amphetamine, on Locomotion, Spatial Cognitive Processing and Neurochemical Profiles in Rats: A Comparison With Immediate-Release Amphetamine (2022) · cites this paper as [11]
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“Therefore, as chronic exposure of the DAT to MA would conceivably predispose the nigrostriatal somatodendritic compartment to continual DA efflux (Khoshbouei et al., 2003; Goodwin et al., 2009), the burden to replenish lost DA stores in the SN may activate signaling mechanisms by which ser31 is increased.”
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“, 2010), and pigeons (Li and McMillan, 2001) and consistent with a shared pharmacological mechanism between methamphetamine and amphetamine (Goodwin et al., 2009).”
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“METH has a stronger effect on DAT-mediated cell physiology than AMPH, whichmay contribute to the euphoric and addictive characteristics of METH compared to amphetamine (AMPH) (Goodwin et al., 2009).”
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Does chronic use of amphetamine-type stimulants impair interference control? – A meta-analysis
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Self-citationPhospho-specific antibodies targeting the amino terminus of the human dopamine transporter
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“Moreover, we and others have demonstrated that phosphorylation of serines in the distal amino (N) terminus of DAT is required for efflux (Fog et al., 2006; Goodwin et al., 2009; Khoshbouei et al., 2004; Ramamoorthy et al., 2011), but not for uptake activity, inhibitor”
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Amphetamine and Methamphetamine Have a Direct and Differential Effect on BV2 Microglia Cells
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“…effect on ChIs, a higher dose of a more potent psychostimulant appears to be required; a single high- September/October 2021, 8(5) ENEURO.0196-21.2021 eNeuro.org dose (30mg/kg) of methamphetamine was found to induce a loss of 29% of ChIs in the dorsal Str (Zhu et al., 2006; Goodwin et al., 2009).”
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“The binding of METH to DAT disrupts DA transport by DAT and enhances the synaptic concentration of DA consequently (Goodwin et al., 2009).”
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“It has been shown that the majority of DAT phosphorylation occurs on one of six N-terminal serine residues (Foster et al., 2002), and that METH stimulates DAT phosphorylation in vitro (Cervinski et al., 2005; Goodwin et al., 2009; Khoshbouei et al., 2004).”
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Psychostimulants and social behaviors
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“Amperometry HEK-YFP-DAT cells were preloaded with DA as described previously (Goodwin et al., 2009; Roth et al., 2013; Sambo et al., 2018).”
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“Goodwin and colleagues (2) have shown that MA induces the release of 5 times more dopamine than amphetamine, which is responsible for the euphoric and addictive properties of MA.”
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“Furthermore, PKC activation has been connected to DAT-mediated DA release, and PKC inhibitors have been shown to block AMPH-mediated DA release in rat striata [48–50].”
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“However, it is possible that these studies are not comparable to the present results based on differences between the effects of METH and D-amphetamine, such as differential dopamine release and DAT modulation [29,63].”
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“…METH (1) interferes with the reuptake of dopamine through the dopamine transporter (DAT), resulting in excessive dopamine in the synaptic cleft (Goodwin et al., 2009); (2) inhibits the storage of dopamine in intracellular vesicles through the vesicular monoamine transporter , thereby…”
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“Methamphetamine is more effective than d-amphetamine in releasing striatal dopamine [30].”
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“Moreover, methamphetamine can also facilitate the non-vesicular release of dopamine via dopamine transporters (Bennett et al., 1998; Goodwin et al., 2009; Xie and Miller, 2009).”
BackgroundAmphetamine und Crystal Meth: Epidemiologie, Symptomatik und Therapie konsumassoziierter Störungen
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“Insge - samt hat Methamphetamin eine stärkere Wirkung auf die Dopaminausschüttung als Amphetamin, was zu den stär-ker euphorisierenden Eigenschaften von Methampheta-min beitragen könnte [19].”
The sleep-wake circuit plays a role in temporal distortions.
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“…methamphetamine appears to generate a more intense and prolonged dopaminergic burden and is associated, particularly under common high-intensity pa tt erns of use, with greater risk of repeated use, dependence, psychosis and neurotoxicity relative to amphetamine [5,7].”
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Metadata
- Authors
- Goodwin, J. S., Larson, G. A., Swant, J., Sen, N., Javitch, J. A., Zahniser, N. R., De Felice, L. J., & Khoshbouei, H.
- Affiliations
- Meharry Medical College (US)
- University of Colorado Denver (US)
- Columbia University (US)
- Corresponding author
- Khoshbouei, Habibeh
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2009-01
- Volume
- 284
- Issue
- 5
- Pages
- 2978-2989
- ISSN
- 0021-9258 (ISSN-L)
- PMC
- PMC2631950
- OpenAlex
- W2025188861
- Cited by
- 203
- References
- 82
- Citations by year
- 2026: 4 · 2025: 14 · 2024: 15 · 2023: 9 · 2022: 19 · 2021: 17 · 2020: 8 · 2019: 10 · 2018: 10 · 2017: 10 · 2016: 12 · 2015: 14 · 2014: 17 · 2013: 6 · 2012: 13
- FWCI
- 3.60
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Foundation (0642188)
- National Institutes of Health (DA012408, DA004216, DA022413)
- National Heart, Lung, and Blood Institute (T32HL076133, HL076133)
- National Institute on Drug Abuse (DA012408, P01DA012408, DA021471, DA015050, K05DA022413, DA-022413, DA-004216, K05DA015050, R01DA026947, DA06338, R24DA021471)
- National Cancer Institute (U54CA091408, U54CA91408)
- National Institute of Neurological Disorders and Stroke (U54NS041071, NS034075, R01NS034075)
- National Center for Research Resources (G12RR003032, U54RR019192, G12-RR03032)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neurotransmitter Receptor Influence on Behavior
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Health Sciences › Medicine › Clinical Neurology › Parkinson's Disease Mechanisms and Treatments
- Keywords
- amphetamine, methamphetamine, dopamine transporter, dopamine release, psychostimulants, intracellular calcium, N-terminal phosphorylation, calcium/calmodulin-dependent protein kinase II, protein kinase C, nucleus accumbens, dopamine clearance, drug addiction
- MeSH terms
- Amphetamine, Animals, Bacterial Proteins, Calcium, Cell Line, Dopamine, Humans, Luminescent Proteins, Male, Methamphetamine, Nucleus Accumbens, Rats, Sprague-Dawley, Patch-Clamp Techniques, Dopamine Plasma Membrane Transport Proteins, Rats, In Vitro Techniques
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