HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 278, Issue 24, 22168-22174, June 13, 2003

This Article Full Text Full Text (PDF) All Versions of this Article: 278/24/22168    most recent M301845200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Loder, M. K. Articles by Melikian, H. E. Search for Related Content PubMed PubMed Citation Articles by Loder, M. K. Articles by Melikian, H. E. Social Bookmarking                      What's this?

The Dopamine Transporter Constitutively Internalizes and Recycles in a Protein Kinase C-regulated Manner in Stably Transfected PC12 Cell Lines^*

Merewyn K. Loder  and Haley E. Melikian   ¶ ||

From the Brudnick Neuropsychiatric Research Institute, Department of Psychiatry, University of Massachusetts School of Medicine, Worcester, Massachusetts 01604, Program in Neuroscience, University of Massachusetts School of Medicine, Worcester, Massachusetts 01604, ^¶ Interdisciplinary Graduate Program, University of Massachusetts School of Medicine, Worcester, Massachusetts 01604

The dopamine transporter (DAT) removes dopamine from the extracellular milieu and is potently inhibited by number of psychoactive drugs, including cocaine, amphetamines, and methylphenidate (Ritalin). Multiple lines of evidence demonstrate that protein kinase C (PKC) down-regulates dopamine transport, primarily by redistributing DAT from the plasma membrane to endosomal compartments, although the mechanisms facilitating transporter sequestration are not defined. Here, we demonstrate that DAT constitutively internalizes and recycles in rat pheochromocytoma (PC12) cells. Temperature blockades demonstrated basal internalization and reliance on recycling to maintain DAT cell surface levels. In contrast, recycling blockade with bafilomycin A₁ significantly decreased transferrin receptor (TfR) surface expression but had no effect on DAT surface levels, suggesting that DAT and TfR traffic via distinct endosomal mechanisms. Kinetic analyses reveal robust constitutive DAT cycling to and from the plasma membrane, independent of transporter expression levels. In contrast, phorbol ester-mediated PKC activation accelerated DAT endocytosis and attenuated transporter recycling in a manner sensitive to DAT expression levels. These data demonstrate constitutive DAT trafficking and that PKC-mediated DAT sequestration is achieved by a combination of accelerated internalization and reduced recycling. Additionally, the differential sensitivity to expression level exhibited by constitutive and regulated DAT trafficking suggests that these two processes are mediated by independent cellular mechanisms.

Received for publication, February 20, 2003 , and in revised form, April 3, 2003.

^* This work was supported by a National Alliance for Research on Schizophrenia and Depression (NARSAD) Young Investigator Award and by National Institutes of Health Grant R01 DA15169-01 (to H. E. M.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| To whom correspondence should be addressed: Brudnick Neuropsychiatric Research Institute, Dept. of Psychiatry, University of Massachusetts Medical School, 303 Belmont St., Worcester, MA 01604. Tel.: 508-856-4045; Fax: 508-856-4130; E-mail: haley.melikian{at}umassmed.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				C. A. Furman, R. Chen, B. Guptaroy, M. Zhang, R. W. Holz, and M. Gnegy Dopamine and Amphetamine Rapidly Increase Dopamine Transporter Trafficking to the Surface: Live-Cell Imaging Using Total Internal Reflection Fluorescence Microscopy J. Neurosci., March 11, 2009; 29(10): 3328 - 3336. [Abstract] [Full Text] [PDF]

				R. Chen, C. A. Furman, M. Zhang, M. N. Kim, R. W. Gereau IV, M. Leitges, and M. E. Gnegy Protein Kinase C{beta} Is a Critical Regulator of Dopamine Transporter Trafficking and Regulates the Behavioral Response to Amphetamine in Mice J. Pharmacol. Exp. Ther., March 1, 2009; 328(3): 912 - 920. [Abstract] [Full Text] [PDF]

				T. Sorkina, T. L. Richards, A. Rao, N. R. Zahniser, and A. Sorkin Negative Regulation of Dopamine Transporter Endocytosis by Membrane-Proximal N-Terminal Residues J. Neurosci., February 4, 2009; 29(5): 1361 - 1374. [Abstract] [Full Text] [PDF]

				S. Li, P. Duan, and G. You Regulation of human organic anion transporter 1 by ANG II: involvement of protein kinase C{alpha} Am J Physiol Endocrinol Metab, February 1, 2009; 296(2): E378 - E383. [Abstract] [Full Text] [PDF]

				Q. Zhang, M. Hong, P. Duan, Z. Pan, J. Ma, and G. You Organic Anion Transporter OAT1 Undergoes Constitutive and Protein Kinase C-regulated Trafficking through a Dynamin- and Clathrin-dependent Pathway J. Biol. Chem., November 21, 2008; 283(47): 32570 - 32579. [Abstract] [Full Text] [PDF]

				M. Pinthong, S. A. G. Black, F. M. Ribeiro, C. Pholpramool, S. S. G. Ferguson, and R. J. Rylett Activity and Subcellular Trafficking of the Sodium-Coupled Choline Transporter CHT Is Regulated Acutely by Peroxynitrite Mol. Pharmacol., March 1, 2008; 73(3): 801 - 812. [Abstract] [Full Text] [PDF]

				P. W. McDonald, S. L. Hardie, T. N. Jessen, L. Carvelli, D. S. Matthies, and R. D. Blakely Vigorous Motor Activity in Caenorhabditis elegans Requires Efficient Clearance of Dopamine Mediated by Synaptic Localization of the Dopamine Transporter DAT-1 J. Neurosci., December 19, 2007; 27(51): 14216 - 14227. [Abstract] [Full Text] [PDF]

				A. Zapata, B. Kivell, Y. Han, J. A. Javitch, E. A. Bolan, D. Kuraguntla, V. Jaligam, M. Oz, L. D. Jayanthi, D. J. Samuvel, et al. Regulation of Dopamine Transporter Function and Cell Surface Expression by D3 Dopamine Receptors J. Biol. Chem., December 7, 2007; 282(49): 35842 - 35854. [Abstract] [Full Text] [PDF]

				M. I. Gonzalez, E. Krizman-Genda, and M. B. Robinson Caveolin-1 Regulates the Delivery and Endocytosis of the Glutamate Transporter, Excitatory Amino Acid Carrier 1 J. Biol. Chem., October 12, 2007; 282(41): 29855 - 29865. [Abstract] [Full Text] [PDF]

				Z. Zhao, X. Li, J. Hao, J. H. Winston, and S. A. Weinman The ClC-3 Chloride Transport Protein Traffics through the Plasma Membrane via Interaction of an N-terminal Dileucine Cluster with Clathrin J. Biol. Chem., September 28, 2007; 282(39): 29022 - 29031. [Abstract] [Full Text] [PDF]

				C.-B. Zhu, J. A. Steiner, J. L. Munn, L. C. Daws, W. A. Hewlett, and R. D. Blakely Rapid Stimulation of Presynaptic Serotonin Transport by A3 Adenosine Receptors J. Pharmacol. Exp. Ther., July 1, 2007; 322(1): 332 - 340. [Abstract] [Full Text] [PDF]

				F. Zhou, M. Hong, and G. You Regulation of human organic anion transporter 4 by progesterone and protein kinase C in human placental BeWo cells Am J Physiol Endocrinol Metab, July 1, 2007; 293(1): E57 - E61. [Abstract] [Full Text] [PDF]

				E. A. Bolan, B. Kivell, V. Jaligam, M. Oz, L. D. Jayanthi, Y. Han, N. Sen, E. Urizar, I. Gomes, L. A. Devi, et al. D2 Receptors Regulate Dopamine Transporter Function via an Extracellular Signal-Regulated Kinases 1 and 2-Dependent and Phosphoinositide 3 Kinase-Independent Mechanism Mol. Pharmacol., May 1, 2007; 71(5): 1222 - 1232. [Abstract] [Full Text] [PDF]

				Y. Liu, J. Dore, and X. Chen Calcium Influx through L-type Channels Generates Protein Kinase M to Induce Burst Firing of Dopamine Cells in the Rat Ventral Tegmental Area J. Biol. Chem., March 23, 2007; 282(12): 8594 - 8603. [Abstract] [Full Text] [PDF]

				S. Urra, C. A. Escudero, P. Ramos, F. Lisbona, E. Allende, P. Covarrubias, J. I. Parraguez, N. Zampieri, M. V. Chao, W. Annaert, et al. TrkA Receptor Activation by Nerve Growth Factor Induces Shedding of the p75 Neurotrophin Receptor Followed by Endosomal {gamma}-Secretase-mediated Release of the p75 Intracellular Domain J. Biol. Chem., March 9, 2007; 282(10): 7606 - 7615. [Abstract] [Full Text] [PDF]

				Y. Wei, J. M. Williams, C. Dipace, U. Sung, J. A. Javitch, A. Galli, and C. Saunders Dopamine Transporter Activity Mediates Amphetamine-Induced Inhibition of Akt through a Ca2+/Calmodulin-Dependent Kinase II-Dependent Mechanism Mol. Pharmacol., March 1, 2007; 71(3): 835 - 842. [Abstract] [Full Text] [PDF]

				M. Miranda, K. R. Dionne, T. Sorkina, and A. Sorkin Three Ubiquitin Conjugation Sites in the Amino Terminus of the Dopamine Transporter Mediate Protein Kinase C-dependent Endocytosis of the Transporter Mol. Biol. Cell, January 1, 2007; 18(1): 313 - 323. [Abstract] [Full Text] [PDF]

				L. D. Jayanthi, B. Annamalai, D. J. Samuvel, U. Gether, and S. Ramamoorthy Phosphorylation of the Norepinephrine Transporter at Threonine 258 and Serine 259 Is Linked to Protein Kinase C-mediated Transporter Internalization J. Biol. Chem., August 18, 2006; 281(33): 23326 - 23340. [Abstract] [Full Text] [PDF]

				J. Idkowiak-Baldys, K. P. Becker, K. Kitatani, and Y. A. Hannun Dynamic Sequestration of the Recycling Compartment by Classical Protein Kinase C J. Biol. Chem., August 4, 2006; 281(31): 22321 - 22331. [Abstract] [Full Text] [PDF]

				T. Sorkina, M. Miranda, K. R. Dionne, B. R. Hoover, N. R. Zahniser, and A. Sorkin RNA Interference Screen Reveals an Essential Role of Nedd4-2 in Dopamine Transporter Ubiquitination and Endocytosis J. Neurosci., August 2, 2006; 26(31): 8195 - 8205. [Abstract] [Full Text] [PDF]

				K. M. Kahlig, B. J. Lute, Y. Wei, C. J. Loland, U. Gether, J. A. Javitch, and A. Galli Regulation of Dopamine Transporter Trafficking by Intracellular Amphetamine Mol. Pharmacol., August 1, 2006; 70(2): 542 - 548. [Abstract] [Full Text] [PDF]

				M. D. Fullerton, L. Wagner, Z. Yuan, and M. Bakovic Impaired trafficking of choline transporter-like protein-1 at plasma membrane and inhibition of choline transport in THP-1 monocyte-derived macrophages Am J Physiol Cell Physiol, April 1, 2006; 290(4): C1230 - C1238. [Abstract] [Full Text] [PDF]

				M. A. Cervinski, J. D. Foster, and R. A. Vaughan Psychoactive Substrates Stimulate Dopamine Transporter Phosphorylation and Down-regulation by Cocaine-sensitive and Protein Kinase C-dependent Mechanisms J. Biol. Chem., December 9, 2005; 280(49): 40442 - 40449. [Abstract] [Full Text] [PDF]

				I. C. Del Castillo, M. Fedor-Chaiken, J. C. Song, V. Starlinger, J. Yoo, K. S. Matlin, and J. B. Matthews Dynamic regulation of Na+-K+-2Cl- cotransporter surface expression by PKC-{epsilon} in Cl--secretory epithelia Am J Physiol Cell Physiol, November 1, 2005; 289(5): C1332 - C1343. [Abstract] [Full Text] [PDF]

				M. Miranda, C. C. Wu, T. Sorkina, D. R. Korstjens, and A. Sorkin Enhanced Ubiquitylation and Accelerated Degradation of the Dopamine Transporter Mediated by Protein Kinase C J. Biol. Chem., October 21, 2005; 280(42): 35617 - 35624. [Abstract] [Full Text] [PDF]

				R. D. Blakely, L. J. DeFelice, and A. Galli Biogenic Amine Neurotransmitter Transporters: Just When You Thought You Knew Them Physiology, August 1, 2005; 20(4): 225 - 231. [Abstract] [Full Text] [PDF]

				B. G. Garcia, Y. Wei, J. A. Moron, R. Z. Lin, J. A. Javitch, and A. Galli Akt Is Essential for Insulin Modulation of Amphetamine-Induced Human Dopamine Transporter Cell-Surface Redistribution Mol. Pharmacol., July 1, 2005; 68(1): 102 - 109. [Abstract] [Full Text] [PDF]

				D. Wang and M. W. Quick Trafficking of the Plasma Membrane {gamma}-Aminobutyric Acid Transporter GAT1: SIZE AND RATES OF AN ACUTELY RECYCLING POOL J. Biol. Chem., May 13, 2005; 280(19): 18703 - 18709. [Abstract] [Full Text] [PDF]

				Y. Haberman, I. Ziv, Y. Gorzalczany, M. Fukuda, and R. Sagi-Eisenberg Classical protein kinase C(s) regulates targeting of synaptotagmin IX to the endocytic recycling compartment J. Cell Sci., April 15, 2005; 118(8): 1641 - 1649. [Abstract] [Full Text] [PDF]

				S. Massari, C. Vanoni, R. Longhi, P. Rosa, and G. Pietrini Protein Kinase C-mediated Phosphorylation of the BGT1 Epithelial {gamma}-Aminobutyric Acid Transporter Regulates Its Association with LIN7 PDZ Proteins: A POST-TRANSLATIONAL MECHANISM REGULATING TRANSPORTER SURFACE DENSITY J. Biol. Chem., February 25, 2005; 280(8): 7388 - 7397. [Abstract] [Full Text] [PDF]

				D. J. Samuvel, L. D. Jayanthi, N. R. Bhat, and S. Ramamoorthy A Role for p38 Mitogen-Activated Protein Kinase in the Regulation of the Serotonin Transporter: Evidence for Distinct Cellular Mechanisms Involved in Transporter Surface Expression J. Neurosci., January 5, 2005; 25(1): 29 - 41. [Abstract] [Full Text] [PDF]

				H. Jiang, Q. Jiang, and J. Feng Parkin Increases Dopamine Uptake by Enhancing the Cell Surface Expression of Dopamine Transporter J. Biol. Chem., December 24, 2004; 279(52): 54380 - 54386. [Abstract] [Full Text] [PDF]

				K. M. Fournier, M. I. Gonzalez, and M. B. Robinson Rapid Trafficking of the Neuronal Glutamate Transporter, EAAC1: EVIDENCE FOR DISTINCT TRAFFICKING PATHWAYS DIFFERENTIALLY REGULATED BY PROTEIN KINASE C AND PLATELET-DERIVED GROWTH FACTOR J. Biol. Chem., August 13, 2004; 279(33): 34505 - 34513. [Abstract] [Full Text] [PDF]

				C. Bjerggaard, J. U. Fog, H. Hastrup, K. Madsen, C. J. Loland, J. A. Javitch, and U. Gether Surface Targeting of the Dopamine Transporter Involves Discrete Epitopes in the Distal C Terminus But Does Not Require Canonical PDZ Domain Interactions J. Neurosci., August 4, 2004; 24(31): 7024 - 7036. [Abstract] [Full Text] [PDF]

				R. A. Vaughan Phosphorylation and Regulation of Psychostimulant-Sensitive Neurotransmitter Transporters J. Pharmacol. Exp. Ther., July 1, 2004; 310(1): 1 - 7. [Abstract] [Full Text] [PDF]

				L.-B. Li, N. Chen, S. Ramamoorthy, L. Chi, X.-N. Cui, L. C. Wang, and M. E. A. Reith The Role of N-Glycosylation in Function and Surface Trafficking of the Human Dopamine Transporter J. Biol. Chem., May 14, 2004; 279(20): 21012 - 21020. [Abstract] [Full Text] [PDF]

				L. D. Jayanthi, D. J. Samuvel, and S. Ramamoorthy Regulated Internalization and Phosphorylation of the Native Norepinephrine Transporter in Response to Phorbol Esters: EVIDENCE FOR LOCALIZATION IN LIPID RAFTS AND LIPID RAFT-MEDIATED INTERNALIZATION J. Biol. Chem., April 30, 2004; 279(18): 19315 - 19326. [Abstract] [Full Text] [PDF]

				K. M. Kahlig, J. A. Javitch, and A. Galli Amphetamine Regulation of Dopamine Transport: COMBINED MEASUREMENTS OF TRANSPORTER CURRENTS AND TRANSPORTER IMAGING SUPPORT THE ENDOCYTOSIS OF AN ACTIVE CARRIER J. Biol. Chem., March 5, 2004; 279(10): 8966 - 8975. [Abstract] [Full Text] [PDF]

				M. B. Robinson Signaling Pathways Take Aim at Neurotransmitter Transporters Sci. Signal., November 4, 2003; 2003(207): pe50 - pe50. [Abstract] [Full Text] [PDF]

				L. Chi and M. E. A. Reith Substrate-Induced Trafficking of the Dopamine Transporter in Heterologously Expressing Cells and in Rat Striatal Synaptosomal Preparations J. Pharmacol. Exp. Ther., November 1, 2003; 307(2): 729 - 736. [Abstract] [Full Text] [PDF]

				J. A. Moron, I. Zakharova, J. V. Ferrer, G. A. Merrill, B. Hope, E. M. Lafer, Z. C. Lin, J. B. Wang, J. A. Javitch, A. Galli, et al. Mitogen-Activated Protein Kinase Regulates Dopamine Transporter Surface Expression and Dopamine Transport Capacity J. Neurosci., September 17, 2003; 23(24): 8480 - 8488. [Abstract] [Full Text] [PDF]