Proteins on this pathway have targeted assays available via the [https://assays.cancer.gov/available_assays?wp_id=WP727 CPTAC Assay Portal] tuberous sclerosis 2 secretory carrier membrane protein 2 retinoblastoma-like 2 (p130) amphiphysin syntaxin 1A (brain) synapsin I poliovirus receptor-related 2 (herpesvirus entry mediator B) tumor necrosis factor solute carrier family 5 (choline transporter), member 7 F-box protein 32 solute carrier family 6 (neurotransmitter transporter, noradrenalin), member 2 integrin, 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-dioxygenase tumor necrosis factor receptor superfamily, member 11b (osteoprotegerin) angiotensinogen (serpin peptidase inhibitor, clade A, member 8) solute carrier family 6 (neurotransmitter transporter, dopamine), member 3 tryptophan hydroxylase 2 histamine receptor H3 mitogen activated protein kinase 14 transforming growth factor beta 1 induced transcript 1 solute carrier family 6 (neurotransmitter transporter, GABA), member 1 interleukin 1 receptor, type I unc-13 homolog B (C. elegans) adenosine A2a receptor tyrosine hydroxylase protein phosphatase 2 (formerly 2A), catalytic subunit, beta isoform interleukin 1 beta solute carrier family 6 (neurotransmitter transporter, serotonin), member 4 Regulation IL1B --+> MAPK14 Western blots showed that IL-1 b and TNFa activated p38 MAPK, in an SB203580-sensitive manner. d77 Mol Transport SLC6A2 ---> noradrenaline 12805287 The norepinephrine transporter (NET) mediates reuptake of norepinephrine released from neurons, and, as such, it is an importantregulator of noradrenergic neurotransmission. 14566944 It will be important in future studies to determine the exactconditions under which cytoplasmic NET is mobilized tothe plasma membrane of PFC NE axons. a55 b83 Mol 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. b9f Mol Transport SLC6A3 ---> dopamine 18614672 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). fc1 Regulation 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). ead Expression AMPH ---| SLC6A3 17032905 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). b9f Regulation 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. a69 Binding SLC6A4 ---- SYN1 18445122 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). bde Regulation IL1R1 ---| IL1B IL-1ra (10 ng/ ml) completely blocked IL-1 b but not anisomycin-induced 5-HT uptake in both (a) midbrain and (b) striatum. d77 Expression AGT --+> SLC6A2 14697674 Angiotensin II increases surface NET expression on brainstem neurons in vitro. 15963952 Depolarization and angiotensin II rapidly increase surface NET expression in primary brainstemcultures [19,20] . cbf a0b Mol 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. a35 Mol 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. c58 Mol Transport ITGB3 ---> serotonin Remark ably, homozygous disruption of Itgb3 led to an approximately 70% decrease in 5-HT uptake (2-tailed, unpaired Student&apos;s t test, P < 0.05, n = 5; Figure 2C). d45 Regulation SLC6A4 ---| MAPK14 16803896 Recent reportsalso show that cell surface levels of SERT can be modified byprotein kinase G (PKG) activation and p38 MAPK inactivation(36). a8f Regulation 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). a8f Regulation reserpine --+> DBH Reserpine selectively increases tyrosine hydroxylase and dopaminebeta-hydroxylase enzyme protein in central noradrenergic neurons. b83 Regulation TNF --+> MAPK14 Western blots showed that IL-1 b and TNFa activated p38 MAPK, in an SB203580-sensitive manner. d77 Expression 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. a35 MolTransport SLC6A4 ---> L-tryptophan Serotonin transporter polymorphism mediates vulnerability to loss of incentivemotivation following acute tryptophan depletion. c84 MolSynthesis 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]. e9b Mol Transport HRH3 ---> noradrenaline 12196911 Activation of histamine H3-receptors inhibits carrier-mediated norepinephrine release during protracted myocardial ischemia. 18674600 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. ca9 d51 MolSynthesis 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. ead MolTransport MAPK14 ---> serotonin 15155839 NECA treatments also lead to activation of p38 MAPK, and p38MAPK inhibitors block NECA stimulation of 5-HT uptake. 16055563 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. cb5 d69 Regulation MAPK14 ---> SLC6A4 18317590 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). d45 Mol Synthesis SLC6A2 ---> dopamine 15103690 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 d5e ProtModification MAPK14 ---> TSC2 Phosphorylation of Cdc25 and tuberin by p38 MAPK enhances interac-tion of these proteins with 14-3-3 [58,59] . cbf Binding SYN1 ---- UNC13B In particular, Munc13 binds to SYN1A and is responsible for phorbol ester-mediated enhancements in neurotransmission ( Betz et al., 1998 ). f47 Type your comment here 17941718 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. 18614672 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. e9b fc1 Mol Transport HEPES ---| SLC5A7 Thus, the possibility exists that HEPES could cause pH-dependent inhibition of hCHT current (Ming et al., 2002). a69 Expression SLC6A2 --+> TH 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. a35 Binding SLC6A4 ---- TGFB1I1 16803896 Associations of SERT with Hic-5 are evident in brain synaptosomes, suggesting the existence of parallel mechanisms operating to regulate SERT at serotonergic synapses. a8f Binding 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. bde MolTransport RBL2 ---> serotonin Results Activation of ARs in RBL-2 H3 Cells Induces an Increase in 5-HT Uptake that Is Enhanced by Sildenafil. cb5 MolSynthesis 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. dab Mol Transport SLC6A2 ---> dopamine 16452680 However, one would expect NE axons to express more plasmalemmal NET to facilitate DA uptake. 15103690 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 14566944 One potentialmechanism may rely on the ability of NET to transport DA(Horn, 1973; Raiteri et al., 1977; Gu et al., 1996). a35 d5e b83 Regulation citalopram ---| SLC6A4 16803896 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). a8f Mol Transport SYN1 ---> epinephrine SYN1A influenced NE transport only in the prese nce of Ca 2+ in brain cortical synaptosomes. f47 Regulation TGFB1I1 ---| SLC6A4 16803896 Hic-5 may compete with otherproteins like MacMARKS that stabilize SERT localization tothe MS leading to SERT inactivation and eventually transporterendocytosis. a8f Expression ACHE --+> SLC5A7 However, AChE / mice did show a dramatic increase in CHT expression. a0f Mol Transport PVRL2 ---> dopamine 18614672 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. fc1 MolTransport IL1B ---> serotonin 16452991 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 d77 Regulation MAPK14 --+> SLC6A4 16452991 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). d77 Binding SLC6A1 ---- STX1A 12629174 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. fa0 MolSynthesis TPH2 ---> serotonin Zhang X, Beaulieu JM, Sotnikova TD, Gainetdinov RR, Caron MG (2004) Tryptophan hydroxylase-2 controls brain serotonin synthesis. b65 Mol Transport SLC6A4 ---> serotonin 15774771 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). 18445122 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. dab bde Regulation STX1A ---| SLC6A1 12629174 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. fa0 Expression MAPK14 --+> SLC6A4 15774771 Theseresults suggest that whereas PKC is involved in regulatedexpression of SERT, p38 MAPK may be involved in main-taining normal/basal expression of SERT dab Mol Transport AMPH ---> noradrenaline AMPH induces NE efflux and may also regulate transporter trafficking. b9f ProtModification MAPK14 ---> CDC25C Phosphorylation of Cdc25 and tuberin by p38 MAPK enhances interac-tion of these proteins with 14-3-3 [58,59] . cbf Mol 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). d77 Regulation AMPH ---> SYN1 Here, we demonstrate that AMPH stimulation of CAMKII stabilizes an hNET/SYN1A complex. b9f Expression MAPK14 --+> TNFRSF11B Inflammatory cytokines activate p38 MAPK to induce osteoprotegerin synthesis by MG-63 cells. d77 Binding SLC6A4 ---- TDO2 19179283 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} b65 MolTransport STX1A ---> noradrenaline Results 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. fa0 Expression TNF --+> FBXO32 TNFalpha acts via p38 MAPK to stimulate expression of the ubiquitin ligase atrogin1/MAFbx in skeletal muscle. d77 Mol Synthesis AGT ---> noradrenaline 14697674 Norepinephrine metabolism in neuronal cultures is increased by angiotensin II.Am. a0b Regulation reserpine --+> TH Reserpine selectively increases tyrosine hydroxylase and dopaminebeta-hydroxylase enzyme protein in central noradrenergic neurons. b83 Mol 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. a69 Binding SCAMP2 ---- SLC6A4 18445122 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 . bde MolTransport MAPK14 --+> SLC6A4 18317590 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). d45 Binding PPP2CB ---- SLC6A4 18445122 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. 15728187 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. bde ead Cell Membrane Outer Membrane Inner Membrane Nucleus monoamine transport pathway PW:0001250 Pathway Ontology regulatory pathway PW:0000004 Pathway Ontology 16452991 PubMed The proinflammatory cytokines interleukin-1beta and tumor necrosis factor-alpha activate serotonin transporters. Neuropsychopharmacology 2006 Zhu CB Blakely RD Hewlett WA 12805287 PubMed A mutation in the human norepinephrine transporter gene (SLC6A2) associated with orthostatic intolerance disrupts surface expression of mutant and wild-type transporters. J Neurosci 2003 Hahn MK Robertson D Blakely RD 14566944 PubMed Ultrastructural 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 Neurol 2003 Miner LH Schroeter S Blakely RD Sesack SR 17032905 PubMed Amphetamine induces a calcium/calmodulin-dependent protein kinase II-dependent reduction in norepinephrine transporter surface expression linked to changes in syntaxin 1A/transporter complexes. Mol Pharmacol 2007 Dipace C Sung U Binda F Blakely RD Galli A 18614672 PubMed Anomalous dopamine release associated with a human dopamine transporter coding variant. J Neurosci 2008 Mazei-Robison MS Bowton E Holy M Schmudermaier M Freissmuth M Sitte HH Galli A Blakely RD 15728187 PubMed p38 MAPK activation elevates serotonin transport activity via a trafficking-independent, protein phosphatase 2A-dependent process. J Biol Chem 2005 Zhu CB Carneiro AM Dostmann WR Hewlett WA Blakely RD 17005849 PubMed Na+, Cl-, and pH dependence of the human choline transporter (hCHT) in Xenopus oocytes: the proton inactivation hypothesis of hCHT in synaptic vesicles. J Neurosci 2006 Iwamoto H Blakely RD De Felice LJ 18445122 PubMed Going with the flow: trafficking-dependent and -independent regulation of serotonin transport. Traffic 2008 Steiner JA Carneiro AM Blakely RD 15963952 PubMed Proteomic analysis of human norepinephrine transporter complexes reveals associations with protein phosphatase 2A anchoring subunit and 14-3-3 proteins. Biochem Biophys Res Commun 2005 Sung U Jennings JL Link AJ Blakely RD 14697674 PubMed Cell surface trafficking of the antidepressant-sensitive norepinephrine transporter revealed with an ectodomain antibody. Mol Cell Neurosci 2003 Savchenko V Sung U Blakely RD 16452680 PubMed Chronic stress increases the plasmalemmal distribution of the norepinephrine transporter and the coexpression of tyrosine hydroxylase in norepinephrine axons in the prefrontal cortex. J Neurosci 2006 Miner LH Jedema HP Moore FW Blakely RD Grace AA Sesack SR 12361670 PubMed Developing novel treatments for mood disorders: accelerating discovery. Biol Psychiatry 2002 Tamminga CA Nemeroff CB Blakely RD Brady L Carter CS Davis KL Dingledine R Gorman JM Grigoriadis DE Henderson DC B Innis RB Killen J Laughren TP McDonald WM M Murphy GM Jr Paul SM Rudorfer MV Sausville E Schatzberg AF Scolnick EM Suppes T 18317590 PubMed Interactions between integrin alphaIIbbeta3 and the serotonin transporter regulate serotonin transport and platelet aggregation in mice and humans. J Clin Invest 2008 Carneiro AM Cook EH Murphy DL Blakely RD 16803896 PubMed Serotonin-, protein kinase C-, and Hic-5-associated redistribution of the platelet serotonin transporter. J Biol Chem 2006 Carneiro AM Blakely RD 17067279 PubMed The functional impact of SLC6 transporter genetic variation. Annu Rev Pharmacol Toxicol 2007 Hahn MK Blakely RD 17941718 PubMed Hypoinsulinemia regulates amphetamine-induced reverse transport of dopamine. PLoS Biol 2007 Williams JM Owens WA Turner GH Saunders C Dipace C Blakely RD France CP Gore JC Daws LC Avison MJ Galli A 12196911 PubMed Monoamine transporter gene structure and polymorphisms in relation to psychiatric and other complex disorders. Pharmacogenomics J 2002 Hahn MK Blakely RD 18674600 PubMed Cholinergic neurons of mouse intrinsic cardiac ganglia contain noradrenergic enzymes, norepinephrine transporters, and the neurotrophin receptors tropomyosin-related kinase A and p75. Neuroscience 2008 Hoard JL Hoover DB Mabe AM Blakely RD Feng N Paolocci N 15155839 PubMed Adenosine receptor, protein kinase G, and p38 mitogen-activated protein kinase-dependent up-regulation of serotonin transporters involves both transporter trafficking and activation. Mol Pharmacol 2004 Zhu CB Hewlett WA Feoktistov I Biaggioni I Blakely RD 16055563 PubMed Human serotonin transporter variants display altered sensitivity to protein kinase G and p38 mitogen-activated protein kinase. Proc Natl Acad Sci U S A 2005 Prasad HC Zhu CB McCauley JL Samuvel DJ Ramamoorthy S Shelton RC Hewlett WA Sutcliffe JS Blakely RD 15103690 PubMed Ultrastructural interactions between terminals expressing the norepinephrine transporter and dopamine neurons in the rat and monkey ventral tegmental area. Synapse 2004 Liprando LA Miner LH Blakely RD Lewis DA Sesack SR 17188889 PubMed Calcium-dependent interactions of the human norepinephrine transporter with syntaxin 1A. Mol Cell Neurosci 2007 Sung U Blakely RD 15774771 PubMed Evidence for biphasic effects of protein kinase C on serotonin transporter function, endocytosis, and phosphorylation. Mol Pharmacol 2005 Jayanthi LD Samuvel DJ Blakely RD Ramamoorthy S 14645660 PubMed Altered striatal function and muscarinic cholinergic receptors in acetylcholinesterase knockout mice. Mol Pharmacol 2003 Volpicelli-Daley LA Hrabovska A Duysen EG Ferguson SM Blakely RD Lockridge O Levey AI 12629174 PubMed A regulated interaction of syntaxin 1A with the antidepressant-sensitive norepinephrine transporter establishes catecholamine clearance capacity. J Neurosci 2003 Sung U Apparsundaram S Galli A Kahlig KM Savchenko V Schroeter S Quick MW Blakely RD 19179283 PubMed Functional coding variation in recombinant inbred mouse lines reveals multiple serotonin transporter-associated phenotypes. Proc Natl Acad Sci U S A 2009 Carneiro AM Airey DC Thompson B Zhu CB Lu L Chesler EJ Erikson KM Blakely RD