5-HT2B receptors are expressed on astrocytes from brain and in culture and are a chronic target for all five conventional ‘serotonin-specific reuptake inhibitors’ | Neuron Glia Biology | Cambridge Core (original) (raw)

Abstract

In well-differentiated primary cultures of mouse astrocytes, which express no serotonin transporter (SERT), the ‘serotonin-specific reuptake inhibitor’ (SSRI) fluoxetine leads acutely to 5-HT2B receptor-mediated, transactivation-dependent phosphorylation of extracellular regulated kinases 1/2 (ERK1/2) with an EC50 of ~5 μM, and chronically to ERK1/2 phosphorylation-dependent upregulation of mRNA and protein expression of calcium-dependent phospholipase A2 (cPLA2) with ten-fold higher affinity. This affinity is high enough that fluoxetine given therapeutically may activate astrocytic 5-HT2B receptors (Li et al., 2008, 2009). We now confirm the expression of 5-HT2B receptors in astrocytes freshly dissociated from mouse brain and isolated by fluorescence-activated cell sorting (FACS) and investigate in cultured cells if the effects of fluoxetine are shared by all five conventional SSRIs with sufficiently high affinity to be relevant for mechanism(s) of action of SSRIs. Phosphorylated and total ERK1/2 and mRNA and protein expression of cPLA2a were determined by Western blot and reverse transcription polymerase chain reaction (RT-PCR). Paroxetine, which differs widely from fluoxetine in affinity for SERT and for another 5-HT2 receptor, the 5-HT2C receptor, acted acutely and chronically like fluoxetine. One micromolar of paroxetine, fluvoxamine or sertraline increased cPLA2a expression during chronic treatment; citalopram had a similar effect at 0.1–0.5 μM; these are therapeutically relevant concentrations.

References

Birkenhager, T.K., van den Broek, W.W., Mulder, P.G., Bruijn, J.A. and Moleman, P. (2004) Comparison of two-phase treatment with imipramine or fluvoxamine, both followed by lithium addition, in inpatients with major depressive disorder. American Journal of Psychiatry 161, 2060–2065.CrossRefGoogle ScholarPubMed

Bjerkenstedt, L., Flyckt, L., Overo, K.F. and Lingjaerde, O. (1985) Relationship between clinical effects, serum drug concentration and serotonin uptake inhibition in depressed patients treated with citalopram. A double-blind comparison of three dose levels. European Journal of Clinical Pharmacology 28, 553–557.CrossRefGoogle ScholarPubMed

Bolden-Watson, C. and Richelson, E. (1993) Blockade by newly developed antidepressants of biogenic amine uptake into rat brain synaptosomes. Life Science 52, 1023–1029.CrossRefGoogle ScholarPubMed

Bolo, N.R., Hode, Y., Nedelec, J.F., Laine, E., Wagner, G. and Macher, J.P. (2000) Brain pharmacokinetics and tissue distribution in vivo of fluvoxamine and fluoxetine by fluorine magnetic resonance spectroscopy. Neuropsychopharmacology 23, 428–438.CrossRefGoogle ScholarPubMed

Bonhaus, D.W., Bach, C., DeSouza, A., Salazar, F.H., Matsuoka, B.D., Zuppan, P. et al. (1995) The pharmacology and distribution of human 5-hydroxytryptamine2B (5-HT2B) receptor gene products: comparison with 5-HT2A and 5-HT2C receptors. British Journal of Pharmacology 115, 622–628.CrossRefGoogle ScholarPubMed

Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248–254.Google ScholarPubMed

Chen, Y., Peng, L., Zhang, X., Stolzenburg, J.U. and Hertz, L. (1995) Further evidence that fluoxetine interacts with a 5-HT2C receptor in glial cells. Brain Research Bulletin 38, 153–159.CrossRefGoogle ScholarPubMed

Deecher, D.C., Wilcox, B.D., Dave, V., Rossman, P.A. and Kimelberg, H.K. (1993) Detection of 5-hydroxytryptamine2 receptors by radioligand binding, northern blot analysis, and Ca2+ responses in rat primary astrocyte cultures. Journal of Neuroscience Research 35, 246–256.CrossRefGoogle ScholarPubMed

DeVane, C.L., Liston, H.L. and Markowitz, J.S. (2002) Clinical pharmacokinetics of sertraline. Clinical Pharmacokinetics 41, 1247–1266.CrossRefGoogle ScholarPubMed

El Marjou, M., Montalescot, V., Buzyn, A. and Geny, B. (2000) Modifications in phospholipase D activity and isoform expression occur upon maturation and differentiation in vivo and in vitro in human myeloid cells. Leukemia 14, 2118–2127.CrossRefGoogle ScholarPubMed

Etienne, N., Schaerlinger, B., Jaffre, F. and Maroteaux, L. (2004) [The 5-HT2B receptor: a main cardio-pulmonary target of serotonin]. Transforming Growth Factor Beta/Physiology 198, 22–29.Google ScholarPubMed

Feng, G., Mellor, R.H., Bernstein, M., Keller-Peck, C., Nguyen, Q.T., Wallace, M. et al. (2000) Imaging neuronal subsets in transgenic mice expressing multiple spectral variants of GFP. Neuron 28, 41–51.CrossRefGoogle ScholarPubMed

Frazer, A. (2001) Serotonergic and noradrenergic reuptake inhibitors: prediction of clinical effects from in vitro potencies. Journal of Clinical Psychiatry 62 Suppl 12, 16–23.Google ScholarPubMed

Freeman, M.P., Nolan, P.E. Jr., Davis, M.F., Anthony, M., Fried, K., Fankhauser, M. et al. (2008) Pharmacokinetics of sertraline across pregnancy and postpartum. Journal of Clinical Psychopharmacology 28, 646–653.CrossRefGoogle ScholarPubMed

Hansson, E., Simonsson, P. and Alling, C. (1987) 5-Hydroxytryptamine stimulates the formation of inositol phosphate in astrocytes from different regions of the brain. Neuropharmacology 26, 1377–1382.CrossRefGoogle ScholarPubMed

Henry, M.E., Schmidt, M.E., Hennen, J., Villafuerte, R.A., Butman, M.L., Tran, P. et al. (2005) A comparison of brain and serum pharmacokinetics of R-fluoxetine and racemic fluoxetine: a 19-F MRS study. Neuropsychopharmacology 30, 1576–1583.CrossRefGoogle ScholarPubMed

Hertz, L., Schousboe, A., Boechler, N., Mukerji, S. and Fedoroff, S. (1978) Kinetic characteristics of the glutamate uptake into normal astrocytes in cultures. Neurochemical Research 3, 1–14.CrossRefGoogle ScholarPubMed

Hertz, L., Bender, A.S., Woodbury, D.M. and White, H.S. (1989) Potassium-stimulated calcium uptake in astrocytes and its potent inhibition by nimodipine. Journal of Neuroscience Research 22, 209–215.CrossRefGoogle ScholarPubMed

Hertz, L., Lovatt, D., Goldman, S.A. and Nedergaard, M. (2010) Adrenoceptors in brain: cellular gene expression and effects on astrocytic metabolism and [Ca(2 + )](i). Neurochemistry International. April 7, Epub ahead of print.CrossRefGoogle Scholar

Hirano, K., Kimura, R., Sugimoto, Y., Yamada, J., Uchida, S., Kato, Y. et al. (2005) Relationship between brain serotonin transporter binding, plasma concentration and behavioural effect of selective serotonin reuptake inhibitors. British Journal of Pharmacology 144, 695–702.CrossRefGoogle ScholarPubMed

Hyttel, J. (1994) Pharmacological characterization of selective serotonin reuptake inhibitors (SSRIs). International Clinical Psychopharmacology 9 supp. 1, 19–26.CrossRefGoogle ScholarPubMed

Hyttel, J., Overo, K.F. and Arnt, J. (1984) Biochemical effects and drug levels in rats after long-term treatment with the specific 5-HT-uptake inhibitor, citalopram. Psychopharmacology (Berlin) 83, 20–27.CrossRefGoogle ScholarPubMed

Ivanov, A.I., Pero, R.S., Scheck, A.C. and Romanovsky, A.A. (2002) Prostaglandin E(2)-synthesizing enzymes in fever: differential transcriptional regulation. American Journal of Physiology. Regulatory, Integrative and Comparative Physiology 283, R1104–R1117.CrossRefGoogle ScholarPubMed

Jusko, W.J. and Gretch, M. (1976) Plasma and tissue protein binding of drugs in pharmacokinetics. Drug Metabolism Reviews 5, 43–140.CrossRefGoogle ScholarPubMed

Kent, J.M., Coplan, J.D., Lombardo, I., Hwang, D.R., Huang, Y., Mawlawi, O. et al. (2002) Occupancy of brain serotonin transporters during treatment with paroxetine in patients with social phobia: a positron emission tomography study with 11C McN 5652. Psychopharmacology (Berlin) 164, 341–348.CrossRefGoogle ScholarPubMed

Kong, E.K., Peng, L., Chen, Y., Yu, A.C. and Hertz, L. (2002) Up-regulation of 5-HT2B receptor density and receptor-mediated glycogenolysis in mouse astrocytes by long-term fluoxetine administration. Neurochemical Research 27, 113–120.CrossRefGoogle ScholarPubMed

Li, Q., Ma, L., Innis, R.B., Seneca, N., Ichise, M., Huang, H. et al. (2004) Pharmacological and genetic characterization of two selective serotonin transporter ligands: 2-[2-(dimethylaminomethylphenylthio)]-5-fluoromethylphenylamine (AFM) and 3-amino-4-[2-(dimethylaminomethyl-phenylthio)]benzonitrile (DASB). Journal of Pharmacology and Experimental Therapeutics 308, 481–486.CrossRefGoogle Scholar

Li, B., Zhang, S., Li, M., Hertz, L. and Peng, L. (2009) Chronic treatment of astrocytes with therapeutically relevant fluoxetine concentrations enhances cPLA2 expression secondary to 5-HT2B-induced, transactivation-mediated ERK1/2 phosphorylation. Psychopharmacology (Berlin) 207, 1–12.CrossRefGoogle ScholarPubMed

Li, B., Zhang, S., Li, M., Hertz, L. and Peng, L. (2010) Serotonin increases ERK(1/2) phosphorylation in astrocytes by stimulation of 5-HT(2B) and 5-HT(2C) receptors. Neurochemistry International (E-Pub.) 5, 2010.Google Scholar

Li, B., Zhang, S., Zhang, H., Nu, W., Cai, L., Hertz, L. et al. (2008) Fluoxetine-mediated 5-HT(2B) receptor stimulation in astrocytes causes EGF receptor transactivation and ERK phosphorylation. Psychopharmacology (Berlin) 201, 443–458.CrossRefGoogle ScholarPubMed

Liu, X., Van Natta, K., Yeo, H., Vilenski, O., Weller, P.E., Worboys, P.D. et al. (2009) Unbound drug concentration in brain homogenate and cerebral spinal fluid at steady state as a surrogate for unbound concentration in brain interstitial fluid. Drug Metabolism and Disposition: The Biological Fate of Chemicals 37, 787–793.CrossRefGoogle ScholarPubMed

Lovatt, D., Sonnewald, U., Waagepetersen, H.S., Schousboe, A., He, W., Lin, J.H. et al. (2007) The transcriptome and metabolic gene signature of protoplasmic astrocytes in the adult murine cortex. The Journal of Neuroscience 27, 12255–12266.CrossRefGoogle ScholarPubMed

Manev, H., Uz, T. and Manev, R. (2003) Glia as a putative target for antidepressant treatments. Journal of Affective Disorders 75, 59–64.CrossRefGoogle ScholarPubMed

Martensson, B., Nyberg, S., Toresson, G., Brodin, E. and Bertilsson, L. (1989) Fluoxetine treatment of depression. Clinical effects, drug concentrations and monoamine metabolites and N-terminally extended substance P in cerebrospinal fluid. Acta Psychiatrica Scandinavica 79, 586–596.CrossRefGoogle ScholarPubMed

Meier, E., Hertz, L. and Schousboe, A. (1991) Neurotransmitters as developmental signals. Neurochemistry International 19, 1–15.CrossRefGoogle Scholar

Meyer, J.H., McMain, S., Kenedy, S.H., Korman, L., Brown, G.M., DaSilva, J.N., Wilson, A.A., Blak, T., Eynan-Harvey, R., Goulding, V.S., Houle, S. and Links, P. (2003) Dysfunctional attitudes and 5-HT2 receptors during depression and self-harm. American Journal of Psychiatry 160, 90–99.CrossRefGoogle ScholarPubMed

Meyer, J.H., Kapur, S., Eisfeld, B., Brown, G.M., Houle, S., DaSilva, J. et al. (2001) The effect of paroxetine on 5-HT(2A) receptors in depression: an [(18)F]setoperone PET imaging study. American Journal of Psychiatry 158, 78–85.CrossRefGoogle Scholar

Meyer, J.H., Wilson, A.A., Sagrati, S., Hussey, D., Carella, A., Potter, W.Z. et al. (2004) Serotonin transporter occupancy of five selective serotonin reuptake inhibitors at different doses: an [11C]DASB positron emission tomography study. American Journal of Psychiatry 161, 826–835.CrossRefGoogle ScholarPubMed

Nilsson, M., Hansson, E. and Ronnback, L. (1991) Adrenergic and 5-HT2 receptors on the same astroglial cell. A microspectrofluorimetric study on cytosolic Ca2+ responses in single cells in primary culture. Brain Research. Developmental Brain Research 63, 33–41.CrossRefGoogle Scholar

Peng, L., Li, B., Du, T., Kong, E.K., Hu, X., Zhang, S. et al. (2010) Astrocytic transactivation by alpha(2)-adrenergic and 5-HT(2B) serotonergic signaling. Neurochemistry International, May 7, Epub ahead of print.CrossRefGoogle Scholar

Pickel, V.M. and Chan, J. (1999) Ultrastructural localization of the serotonin transporter in limbic and motor compartments of the nucleus accumbens. Journal of Neuroscience 19, 7356–7366.CrossRefGoogle ScholarPubMed

Poblete, J.C. and Azmitia, E.C. (1995) Activation of glycogen phosphorylase by serotonin and 3,4-methylenedioxymethamphetamine in astroglial-rich primary cultures: involvement of the 5-HT2A receptor. Brain Research 680, 9–15.CrossRefGoogle Scholar

Rao, J.S., Ertley, R.N., Lee, H.J., Rapoport, S.I. and Bazinet, R.P. (2006) Chronic fluoxetine upregulates activity, protein and mRNA levels of cytosolic phospholipase A2 in rat frontal cortex. Pharmacogenomics Journal 6, 413–420.CrossRefGoogle ScholarPubMed

Reis, M. and Kallen, B. (2010) Delivery outcome after maternal use of antidepressant drugs in pregnancy: an update using Swedish data. Psychological Medicine. Jan. 5, Epub ahead of print.CrossRefGoogle ScholarPubMed

Rhodes, C.J., Davidson, A., Gibbs, J.S., Wharton, J. and Wilkins, M.R. (2009) Therapeutic targets in pulmonary arterial hypertension. Pharmacology and Therapeutics 121, 69–88.CrossRefGoogle ScholarPubMed

Rothman, R.B. and Baumann, M.H. (2009) Serotonergic drugs and valvular heart disease. Expert Opinion on Drug Safety 8, 317–329.CrossRefGoogle ScholarPubMed

Rothman, R.B., Baumann, M.H., Savage, J.E., Rauser, L., McBride, A., Hufeisen, S.J. et al. (2000) Evidence for possible involvement of 5-HT(2B) receptors in the cardiac valvulopathy associated with fenfluramine and other serotonergic medications. Circulation 102, 2836–2841.CrossRefGoogle Scholar

Sanden, N., Thorlin, T., Blomstrand, F., Persson, P.A. and Hansson, E. (2000) 5-Hydroxytryptamine2B receptors stimulate Ca2+ increases in cultured astrocytes from three different brain regions. Neurochemistry International 36, 427–434.CrossRefGoogle ScholarPubMed

Sawamura, K., Suzuki, Y. and Someya, T. (2004) Effects of dosage and CYP2D6-mutated allele on plasma concentration of paroxetine. European Journal of Clinical Pharmacology 60, 553–557.CrossRefGoogle ScholarPubMed

Seki, M., Nawa, H., Morioka, T., Fukuchi, T., Oite, T., Abe, H. et al. (2002) Establishment of a novel enzyme-linked immunosorbent assay for Thy-1; quantitative assessment of neuronal degeneration. Neuroscience Letters 329, 185–188.CrossRefGoogle ScholarPubMed

Sit, D.K., Perel, J.M., Helsel, J.C. and Wisner, K.L. (2008) Changes in antidepressant metabolism and dosing across pregnancy and early postpartum. Journal of Clinical Psychiatry 69, 652–658.CrossRefGoogle ScholarPubMed

Sublette, M.E., Milak, M.S., Hibbeln, J.R., Freed, P.J., Oquendo, M.A., Malone, K.M. et al. (2009) Plasma polyunsaturated fatty acids and regional cerebral glucose metabolism in major depression. Prostaglandins, Leukotrienes and Essential Fatty Acids 80, 57–64.CrossRefGoogle ScholarPubMed

Tatsumi, M., Groshan, K., Blakely, R.D. and Richelson, E. (1997) Pharmacological profile of antidepressants and related compounds at human monoamine transporters. European Journal of Pharmacology 340, 249–258.CrossRefGoogle ScholarPubMed

van Harten, J. (1993) Clinical pharmacokinetics of selective serotonin reuptake inhibitors. Clinical Pharmacokinetics 24, 203–220.CrossRefGoogle ScholarPubMed

Voineskos, A.N., Wilson, A.A., Boovariwala, A., Sagrati, S., Houle, S., Rusjan, P. et al. (2007) Serotonin transporter occupancy of high-dose selective serotonin reuptake inhibitors during major depressive disorder measured with [11C]DASB positron emission tomography. Psychopharmacology (Berlin) 193, 539–545.CrossRefGoogle ScholarPubMed

Weissman, A.M., Levy, B.T., Hartz, A.J., Bentler, S., Donohue, M., Ellingrod, V.L. et al. (2004) Pooled analysis of antidepressant levels in lactating mothers, breast milk, and nursing infants. American Journal of Psychiatry 161, 1066–1078.CrossRefGoogle ScholarPubMed

Wille, S.M., Cooreman, S.G., Neels, H.M. and Lambert, W.E. (2008) Relevant issues in the monitoring and the toxicology of antidepressants. Critical Reviews in Clinical Laboratory Sciences 45, 25–89.CrossRefGoogle ScholarPubMed

Wilson, A.A., Ginovart, N., Hussey, D., Meyer, J. and Houle, S. (2002) In vitro and in vivo characterisation of [11C]-DASB: a probe for in vivo measurements of the serotonin transporter by positron emission tomography. Nuclear Medicine and Biology 29, 509–515.CrossRefGoogle ScholarPubMed

Wong, D.T. and Bymaster, F.P. (1995) Development of antidepressant drugs. Fluoxetine (Prozac) and other selective serotonin uptake inhibitors. Advances in Experimental Medicine and Biology 363, 77–95.CrossRefGoogle ScholarPubMed

Yu, N., Martin, J.L., Stella, N. and Magistretti, P.J. (1993) Arachidonic acid stimulates glucose uptake in cerebral cortical astrocytes. Proceedings of the National Academy of Sciences of the U.S.A. 90, 4042–4046.CrossRefGoogle ScholarPubMed