Abstract
In the central nervous system, serotonergic and dopaminergic signaling is terminated by the activity of specialized transporter proteins for serotonin (SERT) and dopamine (DAT). These transporter proteins are found both on the cell surface and in intracellular transport vesicles. Trafficking between these subcellular domains regulates the efficiency of removal of extracellular neurotransmitters and hence the efficacy of neuronal signaling. Therefore, it is of high interest to gain more insight into the regulatory mechanisms of the human DAT and SERT cell surface expression in their natural surroundings, i.e., in human cells. Because it is not possible to cultivate human neuronal cells expressing these transporter proteins, there is a need to find other human cells expressing these neuronal proteins. Here, we have investigated the expression of human SERT and DAT on developing megakaryocytes and platelet-like particles derived from the megakaryocyte progenitor cell line MEG-01 upon differentiation by valproic acid (VPA) and all-trans retinoic acid (ATRA). Our results show that MEG-01 cells express SERT and DAT and that VPA and ATRA induce a significant increase of transporter expression on developing megakaryocytes and platelets. As compared to ATRA, VPA more efficiently induced SERT expression but not DAT expression. Comparable to naïve platelets and neurons, SERT was localized to both the cell surface and intracellular compartments. Hence, VPA and ATRA-treated MEG-01 cells provide a model well-suited to studying neuronal monoamine transporter expression, not only during transcription and translation but also with respect to protein trafficking to and from the cell surface.
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Ahmed BA, Jeffus BC, Bukhari SI, Harney JT, Unal R, Lupashin VV, van der Sluijs P, Kilic F (2008) Serotonin transamidates Rab4 and facilitates its binding to the C terminus of serotonin transporter. J Biol Chem 283:9388–9398
Ahmed BA, Bukhari IA, Jeffus BC, Harney JT, Thyparambil S, Ziu E, Fraer M, Rusch NJ, Zimniak P, Lupashin V, Tang D, Kilic F (2009) The cellular distribution of serotonin transporter is impeded on serotonin-altered vimentin network. PLoS One 4:e4730
Amara SG, Kuhar MJ (1993) Neurotransmitter transporters: recent progress. Annu Rev Neurosci 16:73–93
Benmansour S, Owens WA, Cecchi M, Morilak DA, Frazer A (2002) Serotonin clearance in vivo is altered to a greater extent by antidepressant-induced downregulation of the serotonin transporter than by acute blockade of this transporter. J Neurosci 22:6766–6772
Brenner B, Harney JT, Ahmed BA, Jeffus BC, Unal R, Mehta JL, Kilic F (2007) Plasma serotonin levels and the platelet serotonin transporter. J Neurochem 102:206–215
Carneiro AM, Blakely RD (2006) Serotonin-, protein kinase C-, and Hic-5-associated redistribution of the platelet serotonin transporter. J Biol Chem 281:24769–24780
Chen NH, Reith ME, Quick MW (2004) Synaptic uptake and beyond: the sodium- and chloride-dependent neurotransmitter transporter family SLC6. Pflugers Arch 447:519–531
Cimino G, Lo-Coco F, Fenu S, Travaglini L, Finolezzi E, Mancini M, Nanni M, Careddu A, Fazi F, Padula F, Fiorini R, Spiriti MA, Petti MC, Venditti A, Amadori S, Mandelli F, Pelicci PG, Nervi C (2006) Sequential valproic acid/all-trans retinoic acid treatment reprograms differentiation in refractory and high-risk acute myeloid leukemia. Cancer Res 66:8903–8911
Coghill D, Banaschewski T (2009) The genetics of attention-deficit/hyperactivity disorder. Expert Rev Neurother 9:1547–1565
Frankhauser P, Grimmer Y, Bugert P, Deuschle M, Schmidt M, Schloss P (2006) Characterization of the neuronal dopamine transporter DAT in human blood platelets. Neurosci Lett 399:197–201
Frankhauser P, Baranyai R, Ahrens T, Schloss P, Deuschle M, Lederbogen F (2008) Platelet surface P-selectin expression is highly correlated with serotonin transporter density in human subjects. Thromb Haemost 100:1201–1203
Giannaccini G, Betti L, Palego L, Schmid L, Fabbrini L, Pelosini C, Gargini C, Da Valle Y, Lanza M, Marsili A, Maffei M, Santini F, Vitti P, Pinchera A, Lucacchini A (2010) Human serotonin transporter expression during megakaryocytic differentiation of MEG-01 cells. Neurochem Res 35:628–635
Haddad PM, Das A, Ashfaq M, Wieck A (2009) A review of valproate in psychiatric practice. Expert Opin Drug Metab Toxicol 5:539–551
Homberg JR, Lesch KP (2011) Looking on the bright side of serotonin transporter gene variation. Biol Psychiatry 69:513–519
Kittler K, Lau T, Schloss P (2010) Antagonists and substrates differentially regulate serotonin transporter cell surface expression in serotonergic neurons. Eur J Pharmacol 629:63–67
Lau T, Horschitz S, Berger S, Bartsch D, Schloss P (2008) Antidepressant-induced internalization of the serotonin transporter in serotonergic neurons. FASEB J 22:1702–1714
Lau T, Schloss P (2011) Differential regulation of serotonin transporter cell surface expression. Wiley Interdiscip Rev on Membrane Transport and Signaling, in press
Lesch KP, Gutknecht L (2005) Pharmacogenetics of the serotonin transporter. Prog Neuropsychopharmacol Biol Psychiatry 29:1062–1073
Lesch KP, Wolozin BL, Murphy DL, Reiderer P (1993) Primary structure of the human platelet serotonin uptake site: identity with the brain serotonin transporter. J Neurochem 60:2319–2322
Lesch KP, Bengel D, Heils A, Sabol SZ, Greenberg BD, Petri S, Benjamin J, Muller CR, Hamer DH, Murphy DL (1996) Association of anxiety-related traits with a polymorphism in the serotonin transporter gene regulatory region. Science 274:1527–1531
Little KY, Elmer LW, Zhong H, Scheys JO, Zhang L (2002) Cocaine induction of dopamine transporter trafficking to the plasma membrane. Mol Pharmacol 61:436–445
Liu WS, Heckman CA (1998) The sevenfold way of PKC regulation. Cell Signal 10:529–542
Mercado CP, Kilic F (2010) Molecular mechanisms of SERT in platelets: regulation of plasma serotonin levels. Mol Interv 10:231–241
Murphy DL, Lerner A, Rudnick G, Lesch KP (2004) Serotonin transporter: gene, genetic disorders, and pharmacogenetics. Mol Interv 4:109–123
Ni W, Watts SW (2006) 5-hydroxytryptamine in the cardiovascular system: focus on the serotonin transporter (SERT). Clin Exp Pharmacol Physiol 33:575–583
Ogura M, Morishima Y, Ohno R, Kato Y, Hirabayashi N, Nagura H, Saito H (1985) Establishment of a novel human megakaryoblastic leukemia cell line, MEG-01, with positive Philadelphia chromosome. Blood 66:1384–1392
Ogura M, Morishima Y, Okumura M, Hotta T, Takamoto S, Ohno R, Hirabayashi N, Nagura H, Saito H (1988) Functional and morphological differentiation induction of a human megakaryoblastic leukemia cell line (MEG-01s) by phorbol diesters. Blood 72:49–60
Pineyro G, Blier P, Dennis T, de Montigny C (1994) Desensitization of the neuronal 5-HT carrier following its long-term blockade. J Neurosci 14:3036–3047
Qian Y, Galli A, Ramamoorthy S, Risso S, DeFelice LJ, Blakely RD (1997) Protein kinase C activation regulates human serotonin transporters in HEK-293 cells via altered cell surface expression. J Neurosci 17:45–57
Schloss P, Puschel AW, Betz H (1994) Neurotransmitter transporters: new members of known families. Curr Opin Cell Biol 6:595–599
Schweinfurth N, Hohmann S, Deuschle M, Lederbogen F, Schloss P (2010) Valproic acid and all trans retinoic acid differentially induce megakaryopoiesis and platelet-like particle formation from the megakaryoblastic cell line MEG-01. Platelets 21:648–657
Steiner JA, Carneiro AM, Blakely RD (2008) Going with the flow: trafficking-dependent and -independent regulation of serotonin transport. Traffic 9:1393–1402
van der Zwaluw CS, Engels RC, Buitelaar J, Verkes RJ, Franke B, Scholte RH (2009) Polymorphisms in the dopamine transporter gene (SLC6A3/DAT1) and alcohol dependence in humans: a systematic review. Pharmacogenomics 10:853–866
Zahniser NR, Doolen S (2001) Chronic and acute regulation of Na+/Cl- -dependent neurotransmitter transporters: drugs, substrates, presynaptic receptors, and signaling systems. Pharmacol Ther 92:21–55
Zahniser NR, Sorkin A (2009) Trafficking of dopamine transporters in psychostimulant actions. Semin Cell Dev Biol 20:411–417
Acknowledgements
This work was supported by the Deutsche Forschungsgemeinschaft (GRK880) to F.L., M.D. and P.S. We thank Dr. Rick Bernardi for critical reading of the manuscript.
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Sarah Hohmann and Nina Schweinfurth have contributed equally to this work
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Hohmann, S., Schweinfurth, N., Lau, T. et al. Differential expression of neuronal dopamine and serotonin transporters DAT and SERT in megakaryocytes and platelets generated from human MEG-01 megakaryoblasts. Cell Tissue Res 346, 151–161 (2011). https://doi.org/10.1007/s00441-011-1256-9
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DOI: https://doi.org/10.1007/s00441-011-1256-9