This pathway describes the process in which neurotransmitters (e.g. monoamines) are moved across cell membranes in the nervous system. Proteins on this pathway have targeted assays available via the [CPTAC Assay Portal](https://assays.cancer.gov/available_assays?wp_id=WP727)tuberous sclerosis 2secretory carrier membrane protein 2retinoblastoma-like 2 (p130)amphiphysinsyntaxin 1A (brain)synapsin Ipoliovirus receptor-related 2 (herpesvirus entry mediator B)tumor necrosis factorsolute carrier family 5 (choline transporter), member 7F-box protein 32solute carrier family 6 (neurotransmitter transporter, noradrenalin), member 2integrin, beta 3 (platelet glycoprotein IIIa, antigen CD61)cell division cycle 25 homolog C (S. pombe)nitric oxide synthase 1 (neuronal)acetylcholinesterase (Yt blood group)dopamine beta-hydroxylase (dopamine beta-monooxygenase)tryptophan 2,3-dioxygenasetumor necrosis factor receptor superfamily, member 11b (osteoprotegerin)angiotensinogen (serpin peptidase inhibitor, clade A, member 8)solute carrier family 6 (neurotransmitter transporter, dopamine), member 3tryptophan hydroxylase 2histamine receptor H3mitogen activated protein kinase 14transforming growth factor beta 1 induced transcript 1solute carrier family 6 (neurotransmitter transporter, GABA), member 1interleukin 1 receptor, type Iunc-13 homolog B (C. elegans)adenosine A2a receptortyrosine hydroxylaseprotein phosphatase 2 (formerly 2A), catalytic subunit, beta isoforminterleukin 1 betasolute carrier family 6 (neurotransmitter transporter, serotonin), member 4Regulation IL1B --+> MAPK14
Western blots showed that IL-1 b and TNFa activated p38 MAPK, in an SB203580-sensitive manner.d77Mol Transport SLC6A2 ---> noradrenaline12805287
The norepinephrine transporter (NET) mediates reuptake of norepinephrine released from neurons, and, as such, it is an importantregulator of noradrenergic neurotransmission.
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It will be important in future studies to determine the exactconditions under which cytoplasmic NET is mobilized tothe plasma membrane of PFC NE axons.a55b83Mol Transport STX1A ---> 4-Aminobutanoic acid
Wang D, Deken SL, Whitworth TL, and Quick MW (2003) Syntaxin 1A inhibits GABA flux, efflux, and exchange mediated by the rat brain GABA transporter GAT1. b9fMol Transport SLC6A3 ---> dopamine18614672
Interestingly,a study examining engineered proline-to-alanine substitutions inTM12 of the rat DAT documented an increased V max for DA efflux (Itokawa et al., 2002). fc1Regulation anisomycin --+> MAPK14
Anisomycin, a protein synthesis inhibitor derived from Streptomyces griseolus , has been shown to trigger p38 MAPK phosphorylation in a variety of cells, including neutro-phils (55), mast cells (37), and myocardial cells (56). eadExpression AMPH ---| SLC6A317032905
In heterologous expression systems and in ex vivo preparations, AMPH exposure diminishes plasma membrane expression of DAT proteins (Saunders et al., 2000; Gulley et al., 2002; Chi and Reith, 2003). b9fRegulation HEPES ---| SLC6A4
Ming L, Farley RA, Lester HA (2002) Voltage-dependent transient currents of human and rat 5-HT transporters (SERT) are blocked by HEPES and ion channel ligands. a69Binding SLC6A4 ---- SYN118445122
Possibly, asSung et al. have proposed for NET, Syn1A associations with SERT dictate whether the transporter contributes to membrane excitability and/or whether the transporterenters a higher capacity 5-HT transport mode (Figure 3). bdeRegulation IL1R1 ---| IL1BIL-1ra (10 ng/ ml) completely blocked IL-1 b but not anisomycin-induced 5-HT uptake in both (a) midbrain and (b) striatum. d77Expression AGT --+> SLC6A214697674
Angiotensin II increases surface NET expression on brainstem neurons in vitro.
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Depolarization and angiotensin II rapidly increase surface NET expression in primary brainstemcultures [19,20] .cbfa0bMol Synthesis TH ---> catecholamines
These observations have always been difficult to understand, given that TH is readily detected in LC cell bodies (Pickel et al., 1975b) and that catecholamine synthesis requires TH.a35Mol Transport ADORA2A ---> serotonin
Mirtazapine is a 2 -receptor antagonist that increases presynaptic release of both serotonin and norepinephrine (de Boer 1996) and is also a 5HT 2 and 5HT 3 antagonist. c58Mol Transport ITGB3 ---> serotonin
Remark ably, homozygous disruption of Itgb3 led to an approximately 70% decrease in 5-HT uptake (2-tailed, unpaired Student's t test, P < 0.05, n = 5; Figure 2C). d45Regulation SLC6A4 ---| MAPK14
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Recent reportsalso show that cell surface levels of SERT can be modified byprotein kinase G (PKG) activation and p38 MAPK inactivation(36). a8fRegulation Phorbol ester --+> SLC6A4
External 5HT can suppress phorbol ester-induced SERT phosphorylation and internalization, providinga mechanism to tune SERT capacity to the demands of 5HTrelease (34).a8fRegulation reserpine --+> DBH
Reserpine selectively increases tyrosine hydroxylase and dopaminebeta-hydroxylase enzyme protein in central noradrenergic neurons. b83Regulation TNF --+> MAPK14
Western blots showed that IL-1 b and TNFa activated p38 MAPK, in an SB203580-sensitive manner. d77Expression TH --+> SLC6A2,16452680 (Tissue: cortex, CellType: neural, CellLineName: NE) If NET and TH localization are linked to activity states, then chronic stress should increase plasmalemmal NET and TH expression, thus altering the dynamics of NE transmission and impacting cognition and affect.
a35MolTransport SLC6A4 ---> L-tryptophan
Serotonin transporter polymorphism mediates vulnerability to loss of incentivemotivation following acute tryptophan depletion. c84MolSynthesis AMPH ---> dopamine
Notably, when examined in rodents, BOLD responses to AMPH are linearly correlated with AMPHinduced changes in extracellular DA levels within thestriatum [54,55]. e9bMol Transport HRH3 ---> noradrenaline12196911
Activation of histamine H3-receptors inhibits carrier-mediated norepinephrine release during protracted myocardial ischemia.
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Hatta E, Yasuda K, Levi R (1997) Activation of histamine H 3 receptors inhibits carrier-mediated norepinephrine release in a human model of protracted myocardial ischemia.ca9d51MolSynthesis anisomycin --+> SB 203580
In the experiments shown, RBL-2H3 cellswere treated with vehicle, NECA, or anisomycin (0.5 and 1.0 M ) for 5 min in KRH assay buffer in the absence/presence of the p38 MAPK inhibitor, SB203580. eadMolTransport MAPK14 ---> serotonin
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NECA treatments also lead to activation of p38 MAPK, and p38MAPK inhibitors block NECA stimulation of 5-HT uptake.
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Altered PKG p38 MAPK sensitivity of 56Ala is evident in native lymphocytes and may involve altered transporter phosphorylation. ( A ) Lymphocytes were genotyped and cultured as described in Materials and Methods and assessed for 5-HT uptake regulation as described for transfected HeLacells.cb5d69Regulation MAPK14 ---> SLC6A4
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Tonic activation p38 MAPK signal ing pathways, without direct phosphorylation of the transporter, controls basal surface SERT expression levels, whereas acute p38 MAPK activation induces trafficking-independent stimulation of SERT transport activity (12, 16). d45Mol Synthesis SLC6A2 ---> dopamine15103690
Given the ability of NET to transport DA (Horn, 1973; Raiteri et al., 1977; Gu et al., 1996), one probable mechanism by which NET inhibitors might increase DA levels is by blocking an important clearance mechanism for DA d5eProtModification MAPK14 ---> TSC2
Phosphorylation of Cdc25 and tuberin by p38 MAPK enhances interac-tion of these proteins with 14-3-3 [58,59] .ccbBinding SYN1 ---- UNC13B
In particular, Munc13 binds to SYN1A and is responsible for phorbol ester-mediated enhancements in neurotransmission ( Betz et al., 1998 ). f4717941718
Here we show that pharmacological manipulation of the PI3K signaling pathway caused by hypoinsulinemic conditions or selective pharmacological inhibition/activation of PI3K dramatically regulates the ability of AMPH to evokeDAT-mediated DA release in the striatum, as determined byHSCA.
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C , D , Representative traces illustrating AMPH evoked DA release in hDAT cells (black) is blocked by 10 M MPH ( C )o r1 0 M cocaine ( D ), whereas 10 M AMPH blocks basal DA efflux in hDAT A559V cells (red) with no further reductions evident with addition of MPH or cocaine. e9bef9Mol Transport HEPES ---| SLC5A7
Thus, the possibility exists that HEPES could cause pH-dependent inhibition of hCHT current (Ming et al., 2002). a69Expression SLC6A2 --+> TH16452680 (Tissue: cortex, CellType: neural, CellLineName: NE) If NET and TH localization are linked to activity states, then chronic stress should increase plasmalemmal NET and TH expression, thus altering the dynamics of NE transmission and impacting cognition and affect.
a35Binding SLC6A4 ---- TGFB1I116803896
Associations of SERT with Hic-5 are evident in brain synaptosomes, suggesting the existence of parallel mechanisms operating to regulate SERT at serotonergic synapses.a8fBinding NOS1 ---- SLC6A4
Evidence of a functional nNOS/SERT association is evident in brain, and synaptosomal 5-HT transport assays using nNOS / mice display increased SERT activity and plasma membrane expression. bdeMolTransport RBL2 ---> serotonin
Results Activation of ARs in RBL-2 H3 Cells Induces an Increase in 5-HT Uptake that Is Enhanced by Sildenafil. cb5MolSynthesis SLC6A4 ---> serotonin
The active transport of 5-HT byplatelet SERT is thought to be important in maintaining thecirculating concentration of 5-HT below the levels required toactivate vascular smooth cells and platelet 5-HT receptors(Stoltz, 1985; Nemeroff et al., 1998; Musselman et al., 2002).Therefore, the above-mentioned findings suggest that SERTfunction in platelets may be regulated by kinases and phos-phatases linked to endogenous receptors and that this regu-lation may play a role in maintaining blood 5-HT levels.dabMol Transport SLC6A2 ---> dopamine16452680
However, one would expect NE axons to express more plasmalemmal NET to facilitate DA uptake.
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Given the ability of NET to transport DA (Horn, 1973; Raiteri et al., 1977; Gu et al., 1996), one probable mechanism by which NET inhibitors might increase DA levels is by blocking an important clearance mechanism for DA
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One potentialmechanism may rely on the ability of NET to transport DA(Horn, 1973; Raiteri et al., 1977; Gu et al., 1996). a35f52b83Regulation citalopram ---| SLC6A416803896
The activation and MS localizationof SERT are inhibited by citalopram, leading to an accumulation of SERTs inthe cytoskeletal-associated fractions and concomitant increase in associa-tions with Hic-5 (CS).a8fMol Transport SYN1 ---> epinephrine
SYN1A influenced NE transport only in the prese nce of Ca 2+ in brain cortical synaptosomes. f47Regulation TGFB1I1 ---| SLC6A416803896
Hic-5 may compete with otherproteins like MacMARKS that stabilize SERT localization tothe MS leading to SERT inactivation and eventually transporterendocytosis.a8fExpression ACHE --+> SLC5A7
However, AChE / mice did show a dramatic increase in CHT expression.a0fMol Transport PVRL2 ---> dopamine18614672
C , D , Representative traces illustrating AMPH evoked DA release in hDAT cells (black) is blocked by 10 M MPH ( C )o r1 0 M cocaine ( D ), whereas 10 M AMPH blocks basal DA efflux in hDAT A559V cells (red) with no further reductions evident with addition of MPH or cocaine.ef9MolTransport IL1B ---> serotonin
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Stimulation of 5-HT Uptake in Mouse Synaptosomes by IL-1 b and TNFa To assess the relevanc e o f o u r findings with cultured raphe cells for native bra in system s, synapto somes were prepared from midbra in and striatu m o f C57B L/6 mice and treat ed with IL-1 b or TNFa
d77Regulation MAPK14 --+> SLC6A4
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We have demonstrated that activation of p38 mitogen-activated protein kinase (MAPK) induces a catalytic activation of the serotonin transporter (SERT) arising from a reduction in the SERT K m for 5-hydroxytryptamine (5-HT). d77Binding SLC6A1 ---- STX1A12629174
Moreover, evidence has been gathered thatsyntaxin 1A forms physical complexes with the NET homologuesGAT1 and GLYT1 (Beckman et al., 1998; Geerlings et al., 2000).In particular, the GAT1 GABA transporters interact with syn-taxin 1A in a phorbol ester-sensitive manner, with evidence thatmultiple PKC-linked receptors can trigger GAT1 redistributionand syntaxin 1A/GAT1 complex disassembly.fa0MolSynthesis TPH2 ---> serotonin
Zhang X, Beaulieu JM, Sotnikova TD, Gainetdinov RR, Caron MG (2004) Tryptophan hydroxylase-2 controls brain serotonin synthesis. b65Mol Transport SLC6A4 ---> serotonin
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Early studies using heterologously expressed hSERTin HEK-293 cells showed that -PMA increased SERT phosphorylation with parallel decreases in the V max for uptake of 5-HT and surface SERT density (Qian et al., 1997; Ramamoorthy et al., 1998a).
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In the absence of nNOS, SERT may remain at the cell surface or target the plasma membrane through a constitutive pathway and thus exhibit theobserved elevations in basal 5-HT transport activity.dabbdeRegulation STX1A ---| SLC6A112629174
Because BoNT/C1 signifi-cantly diminishes GAT1 surface levels, a greater fraction of totalGAT1 proteins may be inhibited constitutively at the plasmamembrane by syntaxin 1A, whereas NET resides mainly intracel-lular, awaiting SNARE-dependent trafficking to the plasmamembrane. fa0Expression MAPK14 --+> SLC6A4
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Theseresults suggest that whereas PKC is involved in regulatedexpression of SERT, p38 MAPK may be involved in main-taining normal/basal expression of SERTdabMol Transport AMPH ---> noradrenaline
AMPH induces NE efflux and may also regulate transporter trafficking.b9fProtModification MAPK14 ---> CDC25C
Phosphorylation of Cdc25 and tuberin by p38 MAPK enhances interac-tion of these proteins with 14-3-3 [58,59] .ccbMol Transport TNF ---> serotonin
RESULTS Stimulation of 5-HT Uptake in RN46A Cells by IL-1 b and TNFa IL-1 b ex erted a concen trationand tim e-dependen t sti mulation of 5-HT trans port activ ity in cultured RN46A cells (Figure 1a and b).d77Regulation AMPH ---> SYN1Here, we demonstrate that AMPH stimulation of CAMKII stabilizes an hNET/SYN1A complex. b9fExpression MAPK14 --+> TNFRSF11B
Inflammatory cytokines activate p38 MAPK to induce osteoprotegerin synthesis by MG-63 cells.d77Binding SLC6A4 ---- TDO2
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Traitsinfluenced by interactions of Slc6a4 and Tph2 variants are prov ided in Table S2 (independent Tph2 -associated traits are pre sented in Table S3 ). CONTEXT{11012153} b65MolTransport STX1A ---> noradrenalineResults NET-mediated NE transport requires syntaxin 1A To evaluate the syntaxin 1A dependence of NET-mediated NEtransport, we established hNET stably transfected CAD cells(CADhNET) and examined whether antisense oligonucleotide-mediated suppression of syntaxin 1A synthesis would influenceNE transport.fa0Expression TNF --+> FBXO32
TNFalpha acts via p38 MAPK to stimulate expression of the ubiquitin ligase atrogin1/MAFbx in skeletal muscle.d77Mol Synthesis AGT ---> noradrenaline14697674
Norepinephrine metabolism in neuronal cultures is increased by angiotensin II.Am.a0bRegulation reserpine --+> TH
Reserpine selectively increases tyrosine hydroxylase and dopaminebeta-hydroxylase enzyme protein in central noradrenergic neurons. b83Mol Transport SLC5A7 ---> Hemicholinium-3
Constitutive leak currents through hCHT depolarize oocytes The selective blocker HC-3 reveals a leak current through hCHT in the absence of choline.a69Binding SCAMP2 ---- SLC6A4
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After confirmation of a native SCAMP2/SERT complex in glutathione S-transferase pull-down assays, these inves-tigators documented a decreased V max of 5-HT transport wh en S CA MP 2 an d S E RT we re c oe x p re s se d in t ra n sf e c te d ce ll s . bdeMolTransport MAPK14 --+> SLC6A4
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Tonic activation p38 MAPK signal ing pathways, without direct phosphorylation of the transporter, controls basal surface SERT expression levels, whereas acute p38 MAPK activation induces trafficking-independent stimulation of SERT transport activity (12, 16). ad9Binding PPP2CB ---- SLC6A4
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The simplest scenario (Figure 3) consistent with the existing data thus involves SERT/PP2Ac associa-tions that are enriched in plasma membrane domains containing active SERT, whereas less active SERT pools and inactivated (and possibly recycling) SERTs beingrelatively deficient in SERT/PP2Ac complexes.
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We speculate that p38 MAPKmay either augment or stabilize SERT/PP2Ac associations, or,alternatively, p38 MAPK may enhance PP2A activity withinmembrane-resident, transporter-phosphatase complexes.bdeeadCell MembraneOuter MembraneInner Membrane19179283PubMedFunctional coding variation in recombinant inbred mouse lines reveals multiple serotonin transporter-associated phenotypes.Proc Natl Acad Sci U S A2009Carneiro AMAirey DCThompson BZhu CBLu LChesler EJErikson KMBlakely RDregulatory pathwayPW:0000004Pathway Ontology18445122PubMedGoing with the flow: trafficking-dependent and -independent regulation of serotonin transport.Traffic2008Steiner JACarneiro AMBlakely RD17067279PubMedThe functional impact of SLC6 transporter genetic variation.Annu Rev Pharmacol Toxicol2007Hahn MKBlakely RD18614672PubMedAnomalous dopamine release associated with a human dopamine transporter coding variant.J Neurosci2008Mazei-Robison MSBowton EHoly MSchmudermaier MFreissmuth MSitte HHGalli ABlakely RD15963952PubMedProteomic analysis of human norepinephrine transporter complexes reveals associations with protein phosphatase 2A anchoring subunit and 14-3-3 proteins.Biochem Biophys Res Commun2005Sung UJennings JLLink AJBlakely RD17005849PubMedNa+, Cl-, and pH dependence of the human choline transporter (hCHT) in Xenopus oocytes: the proton inactivation hypothesis of hCHT in synaptic vesicles.J Neurosci2006Iwamoto HBlakely RDDe Felice LJmonoamine transport pathwayPW:0001250Pathway Ontology15774771PubMedEvidence for biphasic effects of protein kinase C on serotonin transporter function, endocytosis, and phosphorylation.Mol Pharmacol2005Jayanthi LDSamuvel DJBlakely RDRamamoorthy S12361670PubMed12361670PubMedDeveloping novel treatments for mood disorders: accelerating discovery.Biol Psychiatry2002Tamminga CANemeroff CBBlakely RDBrady LCarter CSDavis KLDingledine RGorman JMGrigoriadis DEHenderson DCB Innis RBKillen JLaughren TPMcDonald WMM Murphy GM JrPaul SMRudorfer MVSausville ESchatzberg AFScolnick EMSuppes T16055563PubMedHuman serotonin transporter variants display altered sensitivity to protein kinase G and p38 mitogen-activated protein kinase.Proc Natl Acad Sci U S A2005Prasad HCZhu CBMcCauley JLSamuvel DJRamamoorthy SShelton RCHewlett WASutcliffe JSBlakely RD18674600PubMedCholinergic neurons of mouse intrinsic cardiac ganglia contain noradrenergic enzymes, norepinephrine transporters, and the neurotrophin receptors tropomyosin-related kinase A and p75.Neuroscience2008Hoard JLHoover DBMabe AMBlakely RDFeng NPaolocci N12805287PubMedA mutation in the human norepinephrine transporter gene (SLC6A2) associated with orthostatic intolerance disrupts surface expression of mutant and wild-type transporters.J Neurosci2003Hahn MKRobertson DBlakely RD14697674PubMedCell surface trafficking of the antidepressant-sensitive norepinephrine transporter revealed with an ectodomain antibody.Mol Cell Neurosci2003Savchenko VSung UBlakely RD18317590PubMedInteractions between integrin alphaIIbbeta3 and the serotonin transporter regulate serotonin transport and platelet aggregation in mice and humans.J Clin Invest2008Carneiro AMCook EHMurphy DLBlakely RD17188889PubMedCalcium-dependent interactions of the human norepinephrine transporter with syntaxin 1A.Mol Cell Neurosci2007Sung UBlakely RD12629174PubMedA regulated interaction of syntaxin 1A with the antidepressant-sensitive norepinephrine transporter establishes catecholamine clearance capacity.J Neurosci2003Sung UApparsundaram SGalli AKahlig KMSavchenko VSchroeter SQuick MWBlakely RD16803896PubMedSerotonin-, protein kinase C-, and Hic-5-associated redistribution of the platelet serotonin transporter.J Biol Chem2006Carneiro AMBlakely RD17941718PubMedHypoinsulinemia regulates amphetamine-induced reverse transport of dopamine.PLoS Biol2007Williams JMOwens WATurner GHSaunders CDipace CBlakely RDFrance CPGore JCDaws LCAvison MJGalli A15103690PubMedUltrastructural interactions between terminals expressing the norepinephrine transporter and dopamine neurons in the rat and monkey ventral tegmental area.Synapse2004Liprando LAMiner LHBlakely RDLewis DASesack SR18614672PubMedAnomalous dopamine release associated with a human dopamine transporter coding variant.J Neurosci2008Mazei-Robison MSBowton EHoly MSchmudermaier MFreissmuth MSitte HHGalli ABlakely RD16452991PubMedThe proinflammatory cytokines interleukin-1beta and tumor necrosis factor-alpha activate serotonin transporters.Neuropsychopharmacology2006Zhu CBBlakely RDHewlett WA16452680PubMedChronic stress increases the plasmalemmal distribution of the norepinephrine transporter and the coexpression of tyrosine hydroxylase in norepinephrine axons in the prefrontal cortex.J Neurosci2006Miner LHJedema HPMoore FWBlakely RDGrace AASesack SR12196911PubMedMonoamine transporter gene structure and polymorphisms in relation to psychiatric and other complex disorders.Pharmacogenomics J2002Hahn MKBlakely RD14645660PubMedAltered striatal function and muscarinic cholinergic receptors in acetylcholinesterase knockout mice.Mol Pharmacol2003Volpicelli-Daley LAHrabovska ADuysen EGFerguson SMBlakely RDLockridge OLevey AI15728187PubMedp38 MAPK activation elevates serotonin transport activity via a trafficking-independent, protein phosphatase 2A-dependent process.J Biol Chem2005Zhu CBCarneiro AMDostmann WRHewlett WABlakely RD15155839PubMedAdenosine receptor, protein kinase G, and p38 mitogen-activated protein kinase-dependent up-regulation of serotonin transporters involves both transporter trafficking and activation.Mol Pharmacol2004Zhu CBHewlett WAFeoktistov IBiaggioni IBlakely RD14566944PubMedUltrastructural localization of the norepinephrine transporter in superficial and deep layers of the rat prelimbic prefrontal cortex and its spatial relationship to probable dopamine terminals.J Comp Neurol2003Miner LHSchroeter SBlakely RDSesack SR17032905PubMedAmphetamine induces a calcium/calmodulin-dependent protein kinase II-dependent reduction in norepinephrine transporter surface expression linked to changes in syntaxin 1A/transporter complexes.Mol Pharmacol2007Dipace CSung UBinda FBlakely RDGalli A15103690PubMedUltrastructural interactions between terminals expressing the norepinephrine transporter and dopamine neurons in the rat and monkey ventral tegmental area.Synapse200415103690PubMedUltrastructural interactions between terminals expressing the norepinephrine transporter and dopamine neurons in the rat and monkey ventral tegmental area.Synapse2004Liprando LAMiner LHBlakely RDLewis DASesack SR15963952PubMedProteomic analysis of human norepinephrine transporter complexes reveals associations with protein phosphatase 2A anchoring subunit and 14-3-3 proteins.Biochem Biophys Res Commun200515963952PubMedProteomic analysis of human norepinephrine transporter complexes reveals associations with protein phosphatase 2A anchoring subunit and 14-3-3 proteins.Biochem Biophys Res Commun2005Sung UJennings JLLink AJBlakely RD18317590PubMedInteractions between integrin alphaIIbbeta3 and the serotonin transporter regulate serotonin transport and platelet aggregation in mice and humans.J Clin Invest2008Carneiro AMCook EHMurphy DLBlakely RD