Abstract
(1) Pre-mRNA editing of serotonin 2C (5-HT2C) and glutamate (Glu) receptors (R) influences higher brain functions and pathological states such as epilepsy, amyotrophic lateral sclerosis, and depression. Adenosine deaminases acting on RNA (ADAR1–3) convert adenosine to inosine on synthetic RNAs, analogous to 5-HT2cR and GluR. The order of editing as well as mechanisms controlling editing in native neurons is unknown. (2) With single-cell RT-PCR we investigated the co-expression of ADAR genes with GluR and 5-HT2CR and determined the editing status at known sites in the hypothalamic tuberomamillary nucleus, a major center for wakefulness and arousal. (3) The most frequently expressed enzymes were ADAR1, followed by ADAR2. The Q/R site of GluR2 was always fully edited. Editing at the R/G site in the GluR2 (but not GluR4) subunit was co-ordinated with ADAR expression: maximal editing was found in neurons expressing both ADAR2 splice variants of the deaminase domain and lacking ADAR3. (4) Editing of the 5-HT2CR did not correlate with ADAR expression. The 5-HT2CR mRNA was always edited at A, in the majority of cells at B sites and variably edited at E, C and D sites. A negative correlation was found between editing of C and D sites. The GluR4 R/G site editing was homogeneous within individuals: it was fully edited in all neurons obtained from 12 rats and under-edited in six neurons obtained from three rats. (5) We conclude that GluR2 R/G editing is controlled at the level of ADAR2 and therefore this enzyme may be a target for pharmacotherapy. On the other hand, further factors/enzymes besides ADAR must control or influence 5-HT2CR and GluR pre-mRNA editing in native neurons; our data indicate that these factors vary between individuals and could be predictors of psychiatric disease.
Similar content being viewed by others
References
Audinat E, Lambolez B, Cauli B, Ropert N, Perrais D, Hestrin S, Rossier J (1996) Diversity of glutamate receptors in neocortical neurons: implications for synaptic plasticity. J Physiol Paris 90:331–332
Barbon A, Popoli M, La Via L, Moraschi S, Vallini I, Tardito D, Tiraboschi E, Musazzi L, Giambelli R, Gennarelli M, Racagni G, Barlati S (2006) Regulation of editing and expression of glutamate alpha-amino-propionic-acid (AMPA)/Kainate receptors by antidepressant drugs. Biol Psychiatry 59(8): 713–720
Berg KA, Cropper JD, Niswender CM, Sanders-Bush E, Emeson RB, Clarke WP (2001) RNA-editing of the 5-HT(2C) receptor alters agonist-receptor-effector coupling specificity. Br J Pharmacol 134:386–392
Burns CM, Chu H, Rueter SM, Hutchinson LK, Canton H, Sanders-Bush E, Emeson RB (1997) Regulation of serotonin-2C receptor G-protein coupling by RNA editing. Nature 387:303–308
Chen CX, Cho DS, Wang Q, Lai F, Carter KC, Nishikura K (2000) A third member of the RNA-specific adenosine deaminase gene family, ADAR3, contains both single- and double-stranded RNA binding domains. RNA 6:755–767
Cho DS, Yang W, Lee JT, Shiekhattar R, Murray JM, Nishikura K (2003) Requirement of dimerization for RNA editing activity of adenosine deaminases acting on RNA. J Biol Chem 278(19):17093–17102
Eriksson KS, Sergeeva O, Brown RE, Haas HL (2001a) Orexin/hypocretin excites the histaminergic neurons of the tuberomammillary nucleus. J Neurosci 21:9273–9279
Eriksson KS, Stevens DR, Haas HL (2001b) Serotonin excites tuberomammillary neurons by activation of Na(+)/Ca(2+)-exchange. Neuropharmacology 40:345–351
Eriksson KS, Sergeeva OA, Stevens DR, Haas HL (2002) Neurotransmitter-induced activation of sodium-calcium exchange causes neuronal excitation. Ann N Y Acad Sci 976:405–7:405–407
Esclapez M, Tillakaratne NJ, Kaufman DL, Tobin AJ, Houser CR (1994) Comparative localization of two forms of glutamic acid decarboxylase and their mRNAs in rat brain supports the concept of functional differences between the forms. J Neurosci 14:1834–1855
Gerber A, O’Connell MA, Keller W (1997) Two forms of human double-stranded RNA-specific editase 1 (hRED1) generated by the insertion of an Alu cassette. RNA 3:453–463
Greif KF, Erlander MG, Tillakaratne NJ, Tobin AJ (1991) Postnatal expression of glutamate decarboxylases in developing rat cerebellum. Neurochem Res 16:235–242
Gurevich I, Tamir H, Arango V, Dwork AJ, Mann JJ, Schmauss C (2002a) Altered editing of serotonin 2C receptor pre-mRNA in the prefrontal cortex of depressed suicide victims. Neuron 34:349–356
Gurevich I, Englander MT, Adlersberg M, Siegal NB, Schmauss C (2002b) Modulation of serotonin 2C receptor editing by sustained changes in serotonergic neurotransmission. J Neurosci 22:10529–10532
Hartner JC, Schmittwolf C, Kispert A, Muller AM, Higuchi M, Seeburg PH (2004) Liver disintegration in the mouse embryo caused by deficiency in the RNA-editing enzyme ADAR1. J Biol Chem 279:4894–4902
Higuchi M, Single FN, Kohler M, Sommer B, Sprengel R, Seeburg PH (1993) RNA editing of AMPA receptor subunit GluR-B: a base-paired intron-exon structure determines position and efficiency. Cell 75:1361–1370
Higuchi M, Maas S, Single FN, Hartner J, Rozov A, Burnashev N, Feldmeyer D, Sprengel R, Seeburg PH (2000) Point mutation in an AMPA receptor gene rescues lethality in mice deficient in the RNA-editing enzyme ADAR2. Nature 406:78–81
Kawahara Y, Ito K, Sun H, Aizawa H, Kanazawa I, Kwak S (2004) Glutamate receptors: RNA editing and death of motor neurons. Nature 427:801
Kohr G, Melcher T, Seeburg PH (1998) Candidate editases for GluR channels in single neurons of rat hippocampus and cerebellum. Neuropharmacology 37:1411–1417
Kwak S, Kawahara Y (2005) Deficient RNA editing of GluR2 and neuronal death in amyotropic lateral sclerosis. J Mol Med 83:110–120
Lai F, Chen CX, Carter KC, Nishikura K (1997) Editing of glutamate receptor B subunit ion channel RNAs by four alternatively spliced DRADA2 double-stranded RNA adenosine deaminases. Mol Cell Biol 17:2413–2424
Lambolez B, Audinat E, Bochet P, Crepel F, Rossier J (1992) AMPA receptor subunits expressed by single Purkinje cells. Neuron 9:247–258
Lomeli H, Mosbacher J, Melcher T, Hoger T, Geiger JR, Kuner T, Monyer H, Higuchi M, Bach A, Seeburg PH (1994) Control of kinetic properties of AMPA receptor channels by nuclear RNA editing. Science 266:1709–1713
Maas S, Melcher T, Herb A, Seeburg PH, Keller W, Krause S, Higuchi M, O’Connell MA (1996) Structural requirements for RNA editing in glutamate receptor pre-mRNAs by recombinant double-stranded RNA adenosine deaminase. J Biol Chem 271:12221–12226
Maas S, Patt S, Schrey M, Rich A (2001) Underediting of glutamate receptor GluR-B mRNA in malignant gliomas. Proc Natl Acad Sci U S A 98:14687–14692
Marion S, Weiner DM, Caron MG (2004) RNA editing induces variation in desensitization and trafficking of 5-hydroxytryptamine 2c receptor isoforms. J Biol Chem 279:2945–2954
Melcher T, Maas S, Herb A, Sprengel R, Seeburg PH, Higuchi M (1996a) A mammalian RNA editing enzyme. Nature 379:460–464
Melcher T, Maas S, Herb A, Sprengel R, Higuchi M, Seeburg PH (1996b) RED2, a brain-specific member of the RNA-specific adenosine deaminase family. J Biol Chem 271:31795–31798
Mittaz L, Scott HS, Rossier C, Seeburg PH, Higuchi M, Antonarakis SE (1997) Cloning of a human RNA editing deaminase (ADARB1) of glutamate receptors that maps to chromosome 21q22.3. Genomics 41:210–217
Niswender CM (1998) Recent advances in mammalian RNA editing. Cell Mol Life Sci 54:946–964
Niswender CM, Sanders-Bush E, Emeson RB (1998) Identification and characterization of RNA editing events within the 5-HT2C receptor. Ann N Y Acad Sci 861:38–48
Niswender CM, Copeland SC, Herrick-Davis K, Emeson RB, Sanders-Bush E (1999) RNA editing of the human serotonin 5-hydroxytryptamine 2C receptor silences constitutive activity. J Biol Chem 274:9472–9478
Paul MS, Bass BL (1998) Inosine exists in mRNA at tissue-specific levels and is most abundant in brain mRNA. EMBO J 17:1120–1127
Rueter SM, Dawson TR, Emeson RB (1999) Regulation of alternative splicing by RNA editing. Nature 399:75–80
Schmauss C (2003) Serotonin 2C receptors: suicide, serotonin, and runaway RNA editing. Neuroscientist 9:237–242
Sergeeva OA, Haas HL (2001) Expression and function of glycine receptors in striatal cholinergic interneurons from rat and mouse. Neuroscience 104:1043–1055
Sergeeva OA, Eriksson KS, Sharonova IN, Vorobjev VS, Haas HL (2002) GABA(A) receptor heterogeneity in histaminergic neurons. Eur J Neurosci 16:1472–1482
Sergeeva OA, Amberger BT, Eriksson KS, Scherer A, Haas HL (2003) Coordinated expression of 5-HT2C receptors with the NCX1 Na+/Ca2+ exchanger in histaminergic neurons. J Neurochem 87(3):657–664
Sergeeva OA, Amberger BT, Vorobjev VS, Eriksson KS, Haas HL (2004) AMPA receptor properties and coexpression with sodium-calcium exchangers in rat hypothalamic neurons. Eur J Neurosci 19:957–965
Sergeeva OA, Andreeva N, Garret M, Scherer A, Haas HL (2005a) Pharmacological properties of GABAA receptors in rat hypothalamic neurons expressing the epsilon-subunit. J Neurosci 25:88–95
Sergeeva OA, Schulz D, Doreulee N, Ponomarenko AA, Selbach O, Borsch E, Kircheis G, Huston JP, Haussinger D, Haas HL (2005b) Deficits in cortico-striatal synaptic plasticity and behavioral habituation in rats with portacaval anastomosis. Neuroscience 134:1091–1098
Sodhi MS, Burnet PW, Makoff AJ, Kerwin RW, Harrison PJ (2001) RNA editing of the 5-HT(2C) receptor is reduced in schizophrenia. Mol Psychiatry 6:373–379
Vollmar W, Gloger J, Berger E, Kortenbruck G, Kohling R, Speckmann EJ, Musshoff U (2004) RNA editing (R/G site) and flip-flop splicing of the AMPA receptor subunit GluR2 in nervous tissue of epilepsy patients. Neurobiol Dis 15:371–379
Vorobjev VS, Sharonova IN, Haas HL, Sergeeva OA (2000) Differential modulation of AMPA receptors by cyclothiazide in two types of striatal neurons. Eur J Neurosci 12:2871–2880
Yang JH, Nie Y, Zhao Q, Su Y, Pypaert M, Su H, Rabinovici R (2003) Intracellular localization of differentially regulated RNA-specific adenosine deaminase isoforms in inflammation. J Biol Chem 278:45833–45842
Yang W, Wang Q, Kanes SJ, Murray JM, Nishikura K (2004) Altered RNA editing of serotonin 5-HT2C receptor induced by interferon: implications for depression associated with cytokine therapy. Brain Res Mol Brain Res 124:70–78
Acknowledgments
This study was supported by Deutsche Forschungsgemeinschaft Ha 1525/6–3 and SFB 575. We are very grateful to W. Wisden for discussion and suggestions and to A. Scherer for excellent technical assistance.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sergeeva, O.A., Amberger, B.T. & Haas, H.L. Editing of AMPA and Serotonin 2C Receptors in Individual Central Neurons, Controlling Wakefulness. Cell Mol Neurobiol 27, 669–680 (2007). https://doi.org/10.1007/s10571-007-9153-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10571-007-9153-1