Abstract
Pro-inflammatory cytokines have been implicated in the precipitation of depression and related disorders, and the antidepressant sensitive serotonin transporter (SERT) may be a major target for immune regulation in these disorders. Here, we focus on astrocytes, a major class of immune competent cells in the brain, to examine the effects of pro-longed treatment with tumor necrosis factor-alpha (TNF-α) on SERT activity. We first established that high-affinity serotonin uptake into C6 glioma cells occurs through a SERT-dependent mechanism. Functional SERT expression is also confirmed for primary astrocytes. In both cell types, exposure to TNF-α resulted in a dose- and time-dependent increase in SERT-mediated 5-HT uptake, which was sustained for at least 48 h post-stimulation. Further analysis in primary astrocytes revealed that TNF-α enhanced the transport capacity (Vmax) of SERT-specific 5-HT uptake, suggesting enhanced transporter expression, consistent with our observation of an increase in SERT mRNA levels. We confirmed that in both, primary astrocytes and C6 glioma cells, treatment with TNF-α activates the p38 mitogen-activated protein kinase (MAPK) signaling pathway. Pre-treatment with the p38 MAPK inhibitor SB203580 attenuated the TNF-α mediated stimulation of 5-HT transport in both, C6 glioma and primary astrocytes. In summary, we show that SERT gene expression and activity in astrocytes is subject to regulation by TNF-α, an effect that is at least in part dependent on p38 MAPK activation.
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Ramamoorthy S, Cool DR, Mahesh VB, Leibach FH, Melikian HE, Blakely RD, Ganapathy V (1993) Regulation of the human serotonin transporter. Cholera toxin-induced stimulation of serotonin uptake in human placental choriocarcinoma cells is accompanied by increased serotonin transporter mRNA levels and serotonin transporter-specific ligand binding. J Biol Chem 268(29):21626–21631
Cool DR, Leibach FH, Bhalla VK, Mahesh VB, Ganapathy V (1991) Expression and cyclic AMP-dependent regulation of a high affinity serotonin transporter in the human placental choriocarcinoma cell line (JAR). J Biol Chem 266(24):15750–15757
Rumajogee P, Madeira A, Verge D, Hamon M, Miquel MC (2002) Up-regulation of the neuronal serotoninergic phenotype in vitro: BDNF and cAMP share Trk B-dependent mechanisms. J Neurochem 83(6):1525–1528
Morikawa O, Sakai N, Obara H, Saito N (1998) Effects of interferon-[alpha], interferon-[gamma] and cAMP on the transcriptional regulation of the serotonin transporter. Eur J Pharmacol 349(2–3):317–324
Ramamoorthy JD, Ramamoorthy S, Papapetropoulos A, Catravas JD, Leibach FH, Ganapathy V (1995) Cyclic AMP-independent up-regulation of the human serotonin transporter by staurosporine in choriocarcinoma cells. J Biol Chem 270(29):17189–17195
Bethea CL, Lu NZ, Reddy A, Shlaes T, Streicher JM, Whittemore SR (2003) Characterization of reproductive steroid receptors and response to estrogen in a rat serotonergic cell line. J Neurosci Methods 127(1):31–41
Pecins-Thompson M, Brown NA, Bethea CL (1998) Regulation of serotonin re-uptake transporter mRNA expression by ovarian steroids in rhesus macaques. Brain Res Mol Brain Res 53(1–2):120–129
Glatz K, Mossner R, Heils A, Lesch KP (2003) Glucocorticoid-regulated human serotonin transporter (5-HTT) expression is modulated by the 5-HTT gene-promotor-linked polymorphic region. J Neurochem 86(5):1072–1078
Fumagalli F, Jones SR, Caron MG, Seidler FJ, Slotkin TA (1996) Expression of mRNA coding for the serotonin transporter in aged vs. young rat brain: differential effects of glucocorticoids. Brain Res 719(1–2):225–228
Kubota N, Kiuchi Y, Nemoto M, Oyamada H, Ohno M, Funahashi H, Shioda S, Oguchi K (2001) Regulation of serotonin transporter gene expression in human glial cells by growth factors. Eur J Pharmacol 417(1–2):69–76
Caspi A, Sugden K, Moffitt TE, Taylor A, Craig IW, Harrington H, McClay J, Mill J, Martin J, Braithwaite A, Poulton R (2003) Influence of life stress on depression: moderation by a polymorphism in the 5-HTT gene. Science 301(5631):386–389
Dantzer R, O’Connor JC, Freund GG, Johnson RW, Kelley KW (2008) From inflammation to sickness and depression: when the immune system subjugates the brain. Nat Rev Neurosci 9(1):46–56
Anisman H (2009) Cascading effects of stressors and inflammatory immune system activation: implications for major depressive disorder. J Psychiatry Neurosci JPN 34(1):4–20
Sluzewska A, Rybakowski J, Bosmans E, Sobieska M, Maes M, Wiktorowicz K (1996) Indicators of immune activation in major depression. Psychiatry Res 64(3):161–167
Maes M, Lin A, Delmeire L, Van Gastel A, Kenis G, De Jongh R, Bosmans E (1999) Elevated serum interleukin-6 (IL-6) and IL-6 receptor concentrations in posttraumatic stress disorder following accidental man-made traumatic events. Biol Psychiatry 45(7):833–839
Müller N, Schwarz MJ (2007) The immune-mediated alteration of serotonin and glutamate: towards an integrated view of depression. Mol Psychiatry 12(11):988–1000
Dunn AJ, Swiergiel AH, Beaurepaire R (2005) Cytokines as mediators of depression: what can we learn from animal studies? Neurosci Biobehav Rev 29(4–5):891–909
Schiepers OJG, Wichers MC, Maes M (2005) Cytokines and major depression. Prog Neuropsychopharmacol Biol Psychiatry 29(2):201–217
Simen BB, Duman CH, Simen AA, Duman RS (2006) TNF[alpha] signaling in depression and anxiety: behavioral consequences of individual receptor targeting. Biol Psychiatry 59(9):775–785
Dunn AJ (1992) Endotoxin-induced activation of cerebral catecholamine and serotonin metabolism: comparison with interleukin-1. J Pharmacol Exp Ther 261(3):964–969
Dunn AJ (2006) Effects of cytokines and infections on brain neurochemistry. Clin Neurosci Res 6(1–2):52–68
Kekuda R, Leibach FH, Furesz TC, Smith CH, Ganapathy V (2000) Polarized distribution of interleukin-1 receptors and their role in regulation of serotonin transporter in placenta. J Pharmacol Exp Ther 292(3):1032–1041
Katafuchi T, Kondo T, Take S, Yoshimura M (2005) Enhanced expression of brain interferon-alpha and serotonin transporter in immunologically induced fatigue in rats. Eur J Neurosci 22(11):2817–2826
Ramamoorthy S, Ramamoorthy JD, Prasad PD, Bhat GK, Mahesh VB, Leibach FH, Ganapathy V (1995) Regulation of the human serotonin transporter by interleukin-1[beta]. Biochem Biophys Res Commun 216(2):560–567
Mössner R, Heils A, Stöber G, Okladnova O, Daniel S, Lesch K-P (1998) Enhancement of serotonin transporter function by tumor necrosis factor alpha but not by interleukin-6. Neurochem Int 33(3):251–254
Mössner R, Daniel S, Schmitt A, Albert D, Lesch K-P (2001) Modulation of serotonin transporter function by interleukin-4. Life Sci 68(8):873–880
Zhu C-B, Blakely RD, Hewlett WA (2006) The proinflammatory cytokines interleukin-1beta and tumor necrosis factor-alpha activate serotonin transporters. Neuropsychopharmacology 31(10):2121–2131
Zhu CB, Lindler KM, Owens AW, Daws LC, Blakely RD, Hewlett WA (2010) Interleukin-1 receptor activation by systemic lipopolysaccharide induces behavioral despair linked to MAPK regulation of CNS serotonin transporters. Neuropsychopharmacology 35(13):2510–2520
Hirst WD, Price GW, Rattray M, Wilkin GP (1998) Serotonin transporters in adult rat brain astrocytes revealed by [3H]5-HT uptake into glial plasmalemmal vesicles. Neurochem Int 33(1):11–22
Bel N, Figueras G, Vilaró MT, Suñol C, Artigas F (1997) Antidepressant drugs inhibit a gial 5-hydroxytryptamine transporter in rat brain. Eur J Neurosci 9(8):1728–1738
Katz D, Kimelberg H (1985) Kinetics and autoradiography of high affinity uptake of serotonin by primary astrocyte cultures. J Neurosci 5(7):1901–1908
Dave V, Kimelberg H (1994) Na(+)-dependent, fluoxetine-sensitive serotonin uptake by astrocytes tissue-printed from rat cerebral cortex. J Neurosci 14(8):4972–4986
Inazu M, Takeda H, Ikoshi H, Sugisawa M, Uchida Y, Matsumiya T (2001) Pharmacological characterization and visualization of the glial serotonin transporter. Neurochem Int 39(1):39–49
Pickel VM, Chan J (1999) Ultrastructural localization of the serotonin transporter in limbic and motor compartments of the nucleus accumbens. J Neurosci 19(17):7356–7366
Huang J, Pickel VM (2002) Serotonin transporters (SERTs) within the rat nucleus of the solitary tract: subcellular distribution and relation to 5HT2A receptors. J Neurocytol 31(8–9):667–679
Müller HK, Wiborg O, Haase J (2006) Subcellular redistribution of the serotonin transporter by secretory carrier membrane protein 2. J Biol Chem 281(39):28901–28909
Owens MJ, Morgan WN, Plott SJ, Nemeroff CB (1997) Neurotransmitter receptor and transporter binding profile of antidepressants and their metabolites. J Pharmacol Exp Ther 283(3):1305–1322
Hyttel J (1994) Pharmacological characterization of selective serotonin reuptake inhibitors (SSRIs). Int Clin Psychopharmacol 9(Suppl 1):19–26
Inazu M, Takeda H, Matsumiya T (2003) Expression and functional characterization of the extraneuronal monoamine transporter in normal human astrocytes. J Neurochem 84(1):43–52
Choi S, Friedman WJ (2009) Inflammatory cytokines IL-1beta and TNF-alpha regulate p75NTR expression in CNS neurons and astrocytes by distinct cell-type-specific signalling mechanisms. ASN Neuro 1(2), Article no. e00010. doi:10.1042/AN20090009
Yamaguchi S, Tanabe K, Takai S, Matsushima-Nishiwaki R, Adachi S, Iida H, Kozawa O, Dohi S (2009) Involvement of Rho-kinase in tumor necrosis factor-alpha-induced interleukin-6 release from C6 glioma cells. Neurochem Int 55(6):438–445
Sakai K, Hasegawa C, Okura M, Morikawa O, Ueyama T, Shirai Y, Sakai N, Saito N (2003) Novel variants of murine serotonin transporter mRNA and the promoter activity of its upstream site. Neurosci Lett 342(3):175–178
Foley KF, Pantano C, Ciolino A, Mawe GM (2007) IFN-{gamma} and TNF-{alpha} decrease serotonin transporter function and expression in Caco2 cells. Am J Physiol Gastrointest Liver Physiol 292(3):G779–G784
Huang H, Lung HL, Leung KN, Tsang D (1998) Selective induction of tumor necrosis factor receptor type II gene expression by tumor necrosis factor-[alpha] in C6 glioma cells. Life Sci 62(10):889–896
Samuvel DJ, Jayanthi LD, Bhat NR, Ramamoorthy S (2005) 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 25(1):29–41
Zhu CB, Carneiro AM, Dostmann WR, Hewlett WA, Blakely RD (2005) p38 MAPK activation elevates serotonin transport activity via a trafficking-independent, protein phosphatase 2A-dependent process. J Biol Chem 280(16):15649–15658
Veenstra-VanderWeele J, Muller CL, Iwamoto H, Sauer JE, Owens WA, Shah CR, Cohen J, Mannangatti P, Jessen T, Thompson BJ, Ye R, Kerr TM, Carneiro AM, Crawley JN, Sanders-Bush E, McMahon DG, Ramamoorthy S, Daws LC, Sutcliffe JS, Blakely RD (2012) Autism gene variant causes hyperserotonemia, serotonin receptor hypersensitivity, social impairment and repetitive behavior. Proc Natl Acad Sci USA 109(14):5469–5474
Bruchas MR, Schindler AG, Shankar H, Messinger DI, Miyatake M, Land BB, Lemos JC, Hagan CE, Neumaier JF, Quintana A, Palmiter RD, Chavkin C (2011) Selective p38alpha MAPK deletion in serotonergic neurons produces stress resilience in models of depression and addiction. Neuron 71(3):498–511
Bengel D, Heils A, Petri S, Seemann M, Glatz K, Andrews A, Murphy DL, Lesch KP (1997) Gene structure and 5′-flanking regulatory region of the murine serotonin transporter. Mol Brain Res 44(2):286–292
Nadjar A, Bluthe R-M, May MJ, Dantzer R, Parnet P (2005) Inactivation of the cerebral NF[kappa]B pathway inhibits interleukin-1[beta]-induced sickness behavior and c-Fos expression in various brain nuclei. Neuropsychopharmacology 30(8):1492–1499
Sparacio SM, Zhang Y, Vilcek J, Benveniste EN (1992) Cytokine regulation of interleukin-6 gene expression in astrocytes involves activation of an NF-kappa B-like nuclear protein. J Neuroimmunol 39(3):231–242
Davis RL, Sanchez AC, Lindley DJ, Williams SC, Syapin PJ (2005) Effects of mechanistically distinct NF-kappaB inhibitors on glial inducible nitric-oxide synthase expression. Nitric Oxide Biol Chem Off J Nitric Oxide Soc 12(4):200–209
Fitzgerald LW, Kaplinsky L, Kimelberg H (1990) Serotonin metabolism by monoamine oxidase in rat primary astrocyte cultures. J Neurochem 55(6):2008–2014
Acknowledgments
We thank Dr. Gethin McBean (University College Dublin, Ireland) for the C6 glioma cells. This work was supported by Grants from Science Foundation Ireland (SFI, Grant number BIM0033) and the Irish Health Research Board (HRB, Grant number HRA/2009/303) to J.H.
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Malynn, S., Campos-Torres, A., Moynagh, P. et al. The Pro-inflammatory Cytokine TNF-α Regulates the Activity and Expression of the Serotonin Transporter (SERT) in Astrocytes. Neurochem Res 38, 694–704 (2013). https://doi.org/10.1007/s11064-012-0967-y
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DOI: https://doi.org/10.1007/s11064-012-0967-y