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Disruption of lipid-raft localized Gαs/tubulin complexes by antidepressants: a unique feature of HDAC6 inhibitors, SSRI and tricyclic compounds

Neuropsychopharmacologyvolume 43pages14811491 (2018) | Download Citation

Abstract

Current antidepressant therapies meet with variable therapeutic success and there is increasing interest in therapeutic approaches not based on monoamine signaling. Histone deacetylase 6 (HDAC6), which also deacetylates α-tubulin shows altered expression in mood disorders and HDAC6 knockout mice mimic traditional antidepressant treatments. Nonetheless, a mechanistic understanding for HDAC6 inhibitors in the treatment of depression remains elusive. Previously, we have shown that sustained treatment of rats or glioma cells with several antidepressants translocates Gαs from lipid rafts toward increased association with adenylyl cyclase (AC). Concomitant with this is a sustained increase in cAMP production. While Gαs modifies microtubule dynamics, tubulin also acts as an anchor for Gαs in lipid-rafts. Since HDAC-6 inhibitors potentiate α-tubulin acetylation, we hypothesize that acetylation of α-tubulin disrupts tubulin-Gαs raft-anchoring, rendering Gαs free to activate AC. To test this, C6 Glioma (C6) cells were treated with the HDAC-6 inhibitor, tubastatin-A. Chronic treatment with tubastatin-A not only increased α-tubulin acetylation but also translocated Gαs from lipid-rafts, without changing total Gαs. Reciprocally, depletion of α-tubulin acetyl-transferase-1 ablated this phenomenon. While escitalopram and imipramine also disrupt Gαs/tubulin complexes and translocate Gαs from rafts, they evoke no change in tubulin acetylation. Finally, two indicators of downstream cAMP signaling, cAMP response element binding protein phosphorylation (pCREB) and expression of brain-derived-neurotrophic-factor (BDNF) were both elevated by tubastatin-A. These findings suggest HDAC6 inhibitors show a cellular profile resembling traditional antidepressants, but have a distinct mode of action. They also reinforce the validity of antidepressant-induced Gαs translocation from lipid-rafts as a biosignature for antidepressant response that may be useful in the development of new antidepressant compounds.

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References

  1. 1.

    Covington HE 3rd, Maze I, LaPlant QC, Vialou VF, Ohnishi YN, Berton O, Fass DM, Renthal W, Rush AJ 3rd, Wu EY, Ghose S, Krishnan V, Russo SJ, Tamminga C, Haggarty SJ, Nestler EJ. Antidepressant actions of histone deacetylase inhibitors. J Neurosci. 2009;29:11451–60.

  2. 2.

    Guidotti A, Auta J, Chen Y, Davis JM, Dong E, Gavin DP, Grayson DR, Matrisciano F, Pinna G, Satta R, Sharma RP, Tremolizzo L, Tueting P. Epigenetic GABAergic targets in schizophrenia and bipolar disorder. Neuropharmacology. 2011;60:1007–16.

  3. 3.

    Tsankova N, Renthal W, Kumar A, Nestler EJ. Epigenetic regulation in psychiatric disorders. Nat Rev Neurosci. 2007;8:355–67.

  4. 4.

    Hobara T, Uchida S, Otsuki K, Matsubara T, Funato H, Matsuo K, Suetsugi M, Watanabe Y. Altered gene expression of histone deacetylases in mood disorder patients. J Psychiatr Res. 2010;44:263–70.

  5. 5.

    Russo SJ, Charney DS. Next generation antidepressants. Proc Natl Acad Sci USA. 2013;110:4441–2.

  6. 6.

    Hubbert C, Guardiola A, Shao R, Kawaguchi Y, Ito A, Nixon A, Yoshida M, Wang XF, Yao TP. HDAC6 is a microtubule-associated deacetylase. Nature. 2002;417:455–8.

  7. 7.

    Verdel A, Curtet S, Brocard MP, Rousseaux S, Lemercier C, Yoshida M, Khochbin S. Active maintenance of mHDA2/mHDAC6 histone-deacetylase in the cytoplasm. Curr Biol. 2000;10:747–9.

  8. 8.

    Espallergues J, Teegarden SL, Veerakumar A, Boulden J, Challis C, Jochems J, Chan M, Petersen T, Deneris E, Matthias P, Hahn CG, Lucki I, Beck SG, Berton O. HDAC6 regulates glucocorticoid receptor signaling in serotonin pathways with critical impact on stress resilience. J Neurosci. 2012;32:4400–16.

  9. 9.

    Jochems J, Boulden J, Lee BG, Blendy JA, Jarpe M, Mazitschek R, Van Duzer JH, Jones S, Berton O. Antidepressant-like properties of novel HDAC6-selective inhibitors with improved brain bioavailability. Neuropsychopharmacology. 2014;39:389–400.

  10. 10.

    Lee JB, Wei J, Liu W, Cheng J, Feng J, Yan Z. Histone deacetylase 6 gates the synaptic action of acute stress in prefrontal cortex. J Physiol. 2012;590:1535–46.

  11. 11.

    Fukada M, Hanai A, Nakayama A, Suzuki T, Miyata N, Rodriguiz RM, Wetsel WC, Yao TP, Kawaguchi Y. Loss of deacetylation activity of Hdac6 affects emotional behavior in mice. PLoS ONE. 2012;7:e30924.

  12. 12.

    Bianchi M, Baulieu EE. 3beta-Methoxy-pregnenolone (MAP4343) as an innovative therapeutic approach for depressive disorders. Proc Natl Acad Sci USA. 2012;109:1713–8.

  13. 13.

    Bianchi M, Hagan JJ, Heidbreder CA. Neuronal plasticity, stress and depression: involvement of the cytoskeletal microtubular system? Curr Drug Targets CNS Neurol Disord. 2005;4:597–611.

  14. 14.

    Bianchi M, Heidbreder C, Crespi F. Cytoskeletal changes in the hippocampus following restraint stress: role of serotonin and microtubules. Synapse. 2003;49:188–94.

  15. 15.

    Duman RS, Malberg J, Nakagawa S, D’Sa C. Neuronal plasticity and survival in mood disorders. Biol Psychiatry. 2000;48:732–9.

  16. 16.

    Carlier MF, Didry D, Valentin-Ranc C. Interaction between chromium GTP and tubulin. Stereochemistry of GTP binding, GTP hydrolysis, and microtubule stabilization. J Biol Chem. 1991;266:12361–8.

  17. 17.

    Valenzuela-Fernandez A, Cabrero JR, Serrador JM, Sanchez-Madrid F. HDAC6: a key regulator of cytoskeleton, cell migration and cell-cell interactions. Trends Cell Biol. 2008;18:291–7.

  18. 18.

    Zhang Y, Li N, Caron C, Matthias G, Hess D, Khochbin S, Matthias P. HDAC-6 interacts with and deacetylates tubulin and microtubules in vivo. EMBO J. 2003;22:1168–79.

  19. 19.

    Takemura R, Okabe S, Umeyama T, Kanai Y, Cowan NJ, Hirokawa N. Increased microtubule stability and alpha tubulin acetylation in cells transfected with microtubule-associated proteins MAP1B, MAP2 or tau. J Cell Sci. 1992;103:953–64. (Pt 4)

  20. 20.

    Rivieccio MA, Brochier C, Willis DE, Walker BA, D’Annibale MA, McLaughlin K, Siddiq A, Kozikowski AP, Jaffrey SR, Twiss JL, Ratan RR, Langley B. HDAC6 is a target for protection and regeneration following injury in the nervous system. Proc Natl Acad Sci USA. 2009;106:19599–604.

  21. 21.

    Creppe C, Malinouskaya L, Volvert ML, Gillard M, Close P, Malaise O, Laguesse S, Cornez I, Rahmouni S, Ormenese S, Belachew S, Malgrange B, Chapelle JP, Siebenlist U, Moonen G, Chariot A, Nguyen L. Elongator controls the migration and differentiation of cortical neurons through acetylation of alpha-tubulin. Cell. 2009;136:551–64.

  22. 22.

    Fujita M, Hines CS, Zoghbi SS, Mallinger AG, Dickstein LP, Liow JS, Zhang Y, Pike VW, Drevets WC, Innis RB, Zarate CA Jr.. Downregulation of brain phosphodiesterase type IV measured with 11C-(R)-rolipram positron emission tomography in major depressive disorder. Biol Psychiatry. 2012;72:548–54.

  23. 23.

    Hines LM, Tabakoff B, W.I.S.o. State, U. Trait Markers of Alcohol, and I. Dependence. Platelet adenylyl cyclase activity: a biological marker for major depression and recent drug use. Biol Psychiatry. 2005;58:955–62.

  24. 24.

    Mooney JJ, Samson JA, McHale NL, Pappalarado KM, Alpert JE, Schildkraut JJ. Increased Gsalpha within blood cell membrane lipid microdomains in some depressive disorders: an exploratory study. J Psychiatr Res. 2013;47:706–11.

  25. 25.

    Fujita M, Richards EM, Niciu MJ, Ionescu DF, Zoghbi SS, Hong J, Telu S, Hines CS, Pike VW, Zarate CA, Innis RB. cAMP signaling in brain is decreased in unmedicated depressed patients and increased by treatment with a selective serotonin reuptake inhibitor. Mol Psychiatry. 2017;22:754–9.

  26. 26.

    Allen JA, Halverson-Tamboli RA, Rasenick MM. Lipid raft microdomains and neurotransmitter signalling. Nat Rev Neurosci. 2007;8:128–40.

  27. 27.

    Brown DA. Lipid rafts, detergent-resistant membranes, and raft targeting signals. Physiology. 2006;21:430–9.

  28. 28.

    Allen JA, Yu JZ, Dave RH, Bhatnagar A, Roth BL, Rasenick MM. Caveolin-1 and lipid microdomains regulate Gs trafficking and attenuate Gs/adenylyl cyclase signaling. Mol Pharmacol. 2009;76:1082–93.

  29. 29.

    Donati RJ, Rasenick MM. Chronic antidepressant treatment prevents accumulation of gsalpha in cholesterol-rich, cytoskeletal-associated, plasma membrane domains (lipid rafts). Neuropsychopharmacology . 2005;30:1238–45.

  30. 30.

    Allen JA, Yu JZ, Donati RJ, Rasenick MM. Beta-adrenergic receptor stimulation promotes G alpha s internalization through lipid rafts: a study in living cells. Mol Pharmacol. 2005;67:1493–504.

  31. 31.

    Insel PA, Head BP, Ostrom RS, Patel HH, Swaney JS, Tang CM, Roth DM. Caveolae and lipid rafts: G protein-coupled receptor signaling microdomains in cardiac myocytes. Ann N Y Acad Sci. 2005a;1047:166–72.

  32. 32.

    Insel PA, Head BP, Patel HH, Roth DM, Bundey RA, Swaney JS. Compartmentation of G-protein-coupled receptors and their signalling components in lipid rafts and caveolae. Biochem Soc Trans. 2005b;33:1131–4.

  33. 33.

    Bayewitch ML, Nevo I, Avidor-Reiss T, Levy R, Simonds WF, Vogel Z. Alterations in detergent solubility of heterotrimeric G proteins after chronic activation of G(i/o)-coupled receptors: changes in detergent solubility are in correlation with onset of adenylyl cyclase superactivation. Mol Pharmacol. 2000;57:820–5.

  34. 34.

    Moffett S, Brown DA, Linder ME. Lipid-dependent targeting of G proteins into rafts. J Biol Chem. 2000;275:2191–8.

  35. 35.

    Ostrom RS, Insel PA. The evolving role of lipid rafts and caveolae in G protein-coupled receptor signaling: implications for molecular pharmacology. Br J Pharmacol. 2004;143:235–45.

  36. 36.

    Rybin VO, Xu X, Lisanti MP, Steinberg SF. Differential targeting of beta -adrenergic receptor subtypes and adenylyl cyclase to cardiomyocyte caveolae. A mechanism to functionally regulate the cAMP signaling pathway. J Biol Chem. 2000;275:41447–57.

  37. 37.

    Donati RJ, Dwivedi Y, Roberts RC, Conley RR, Pandey GN, Rasenick MM. Postmortem brain tissue of depressed suicides reveals increased Gs alpha localization in lipid raft domains where it is less likely to activate adenylyl cyclase. J Neurosci. 2008;28:3042–50.

  38. 38.

    Toki S, Donati RJ, Rasenick MM. Treatment of C6 glioma cells and rats with antidepressant drugs increases the detergent extraction of G(s alpha) from plasma membrane. J Neurochem. 1999;73:1114–20.

  39. 39.

    Chen J, Rasenick MM. Chronic treatment of C6 glioma cells with antidepressant drugs increases functional coupling between a G protein (Gs) and adenylyl cyclase. J Neurochem. 1995;64:724–32.

  40. 40.

    Erb SJ, Schappi JM, Rasenick MM. Antidepressants accumulate in lipid rafts independent of monoamine transporters to modulate redistribution of the G protein, Galphas. J Biol Chem. 2016;291:19725–33.

  41. 41.

    Czysz AH, Schappi JM, Rasenick MM. Lateral diffusion of Galphas in the plasma membrane is decreased after chronic but not acute antidepressant treatment: role of lipid raft and non-raft membrane microdomains. Neuropsychopharmacology. 2015;40:766–73.

  42. 42.

    Nelson JC, Thase ME, Trivedi MH, Fava M, Han J, Van Tran Q, et al. Safety and Tolerability of Adjunctive Aripiprazole in Major Depressive Disorder: A Pooled Post Hoc Analysis (studies CN138-139 and CN138-163). Prim. Care Companion J Clin Psychiatry. 2009

  43. 43.

    Zhang L, Rasenick MM. Chronic treatment with escitalopram but not R-citalopram translocates Galpha(s) from lipid raft domains and potentiates adenylyl cyclase: a 5-hydroxytryptamine transporter-independent action of this antidepressant compound. J Pharmacol Exp Ther. 2010;332:977–84.

  44. 44.

    Head BP, Patel HH, Roth DM, Murray F, Swaney JS, Niesman IR, Farquhar MG, Insel PA. Microtubules and actin microfilaments regulate lipid raft/caveolae localization of adenylyl cyclase signaling components. J Biol Chem. 2006;281:26391–9.

  45. 45.

    Dave RH, Saengsawang W, Yu JZ, Donati R, Rasenick MM. Heterotrimeric G-proteins interact directly with cytoskeletal components to modify microtubule-dependent cellular processes. Neurosignals. 2009;17:100–8.

  46. 46.

    Layden BT, Saengsawang W, Donati RJ, Yang S, Mulhearn DC, Johnson ME, Rasenick MM. Structural model of a complex between the heterotrimeric G protein, Gsalpha, and tubulin. Biochim Biophys Acta. 2008;1783:964–73.

  47. 47.

    Nogales E, Wolf SG, Downing KH. Structure of the alpha beta tubulin dimer by electron crystallography. Nature. 1998;391:199–203.

  48. 48.

    Takano K, Yamasaki H, Kawabe K, Moriyama M, Nakamura Y. Imipramine induces brain-derived neurotrophic factor mRNA expression in cultured astrocytes. J Pharmacol Sci. 2012;120:176–86.

  49. 49.

    Eshleman AJ, Stewart E, Evenson AK, Mason JN, Blakely RD, Janowsky A, Neve KA. Metabolism of catecholamines by catechol-O-methyltransferase in cells expressing recombinant catecholamine transporters. J Neurochem. 1997;69:1459–66.

  50. 50.

    Sanacora G, Banasr M. From pathophysiology to novel antidepressant drugs: glial contributions to the pathology and treatment of mood disorders. Biol Psychiatry. 2013;73:1172–9.

  51. 51.

    Manev H, Uz T, Manev R. Glia as a putative target for antidepressant treatments. J Affect Disord. 2003;75:59–64.

  52. 52.

    Schipke CG, Heuser I, Peters O. Antidepressants act on glial cells: SSRIs and serotonin elicit astrocyte calcium signaling in the mouse prefrontal cortex. J Psychiatr Res. 2011;45:242–8.

  53. 53.

    Golan M, Schreiber G, Avissar S. Antidepressants elevate GDNF expression and release from C(6) glioma cells in a beta-arrestin1-dependent, CREB interactive pathway. Int J Neuropsychopharmacol. 2011;14:1289–1300.

  54. 54.

    Hisaoka K, Maeda N, Tsuchioka M, Takebayashi M. Antidepressants induce acute CREB phosphorylation and CRE-mediated gene expression in glial cells: a possible contribution to GDNF production. Brain Res. 2008;1196:53–58.

  55. 55.

    Zhang Y, Kwon S, Yamaguchi T, Cubizolles F, Rousseaux S, Kneissel M, Cao C, Li N, Cheng HL, Chua K, Lombard D, Mizeracki A, Matthias G, Alt FW, Khochbin S, Matthias P. Mice lacking histone deacetylase 6 have hyperacetylated tubulin but are viable and develop normally. Mol Cell Biol. 2008;28:1688–701.

  56. 56.

    Popoli M, Brunello N, Perez J, Racagni G. Second messenger-regulated protein kinases in the brain: their functional role and the action of antidepressant drugs. J Neurochem. 2000;74:21–33.

  57. 57.

    Kalil K, Szebenyi G, Dent EW. Common mechanisms underlying growth cone guidance and axon branching. J Neurobiol. 2000;44:145–58.

  58. 58.

    Menninger JA, Tabakoff B. Forskolin-stimulated platelet adenylyl cyclase activity is lower in persons with major depression. Biol Psychiatry. 1997;42:30–38.

  59. 59.

    Rochlin MW, Wickline KM, Bridgman PC. Microtubule stability decreases axon elongation but not axoplasm production. J Neurosci. 1996;16:3236–46.

  60. 60.

    Scifo E, Pabba M, Kapadia F, Ma T, Lewis DA, Tseng GC, et al. Sustained Molecular Pathology Across Episodes and Remission in Major Depressive Disorder. Biol Psychiatry. 2017

  61. 61.

    Bianchi M, Shah AJ, Fone KC, Atkins AR, Dawson LA, Heidbreder CA, Hows ME, Hagan JJ, Marsden CA. Fluoxetine administration modulates the cytoskeletal microtubular system in the rat hippocampus. Synapse. 2009b;63:359–64.

  62. 62.

    Bianchi M, Fone KC, Shah AJ, Atkins AR, Dawson LA, Heidbreder CA, Hagan JJ, Marsden CA. Chronic fluoxetine differentially modulates the hippocampal microtubular and serotonergic system in grouped and isolation reared rats. Eur Neuropsychopharmacol. 2009a;19:778–90.

  63. 63.

    Bianchi, M., Shah, A.J. Fone, K.C. Atkins, A.R. Dawson, L.A. Heidbreder, C.A. et al. Fluoxetine administration modulates the cytoskeletal microtubular system in the rat hippocampus. Synapse. 2009b.

  64. 64.

    Ladurelle N, Gabriel C, Viggiano A, Mocaer E, Baulieu EE, Bianchi M. Agomelatine (S20098) modulates the expression of cytoskeletal microtubular proteins, synaptic markers and BDNF in the rat hippocampus, amygdala and PFC. Psychopharmacology. 2012;221:493–509.

  65. 65.

    Tsvetanova NG, von Zastrow M. Spatial encoding of cyclic AMP signaling specificity by GPCR endocytosis. Nat Chem Biol. 2014;10:1061–5.

  66. 66.

    Sarma T, Koutsouris A, Yu JZ, Krbanjevic A, Hope TJ, Rasenick MM. Activation of microtubule dynamics increases neuronal growth via the nerve growth factor (NGF)- and Galphas-mediated signaling pathways. J Biol Chem. 2015;290:10045–56.

  67. 67.

    Yu JZ, Dave RH, Allen JA, Sarma T, Rasenick MM. Cytosolic G{alpha}s acts as an intracellular messenger to increase microtubule dynamics and promote neurite outgrowth. J Biol Chem. 2009;284:10462–72.

  68. 68.

    Dave RH, Saengsawang W, Lopus M, Dave S, Wilson L, Rasenick MM. A molecular and structural mechanism for G protein-mediated microtubule destabilization. J Biol Chem. 2011;286:4319–28.

  69. 69.

    Piperno G, LeDizet M, Chang XJ. Microtubules containing acetylated alpha-tubulin in mammalian cells in culture. J Cell Biol. 1987;104:289–302.

  70. 70.

    Webster DR, Borisy GG. Microtubules are acetylated in domains that turn over slowly. J Cell Sci. 1989;92:57–65. (Pt 1)

  71. 71.

    Yu H, Wakim B, Li M, Halligan B, Tint GS, Patel SB. Quantifying raft proteins in neonatal mouse brain by ‘tube-gel’ protein digestion label-free shotgun proteomics. Proteome Sci. 2007;5:17.

  72. 72.

    Menkes DB, Rasenick MM, Wheeler MA, Bitensky MW. Guanosine triphosphate activation of brain adenylate cyclase: enhancement by long-term antidepressant treatment. Science. 1983;219:65–67.

  73. 73.

    Blendy JA. The role of CREB in depression and antidepressant treatment. Biol Psychiatry. 2006;59:1144–50.

  74. 74.

    Nair A, Vaidya VA. Cyclic AMP response element binding protein and brain-derived neurotrophic factor: molecules that modulate our mood? J Biosci. 2006;31:423–34.

  75. 75.

    Nibuya M, Nestler EJ, Duman RS. Chronic antidepressant administration increases the expression of cAMP response element binding protein (CREB) in rat hippocampus. J Neurosci. 1996;16:2365–72.

  76. 76.

    Conti AC, Cryan JF, Dalvi A, Lucki I, Blendy JA. cAMP response element-binding protein is essential for the upregulation of brain-derived neurotrophic factor transcription, but not the behavioral or endocrine responses to antidepressant drugs. J Neurosci. 2002;22:3262–8.

  77. 77.

    Nibuya M, Morinobu S, Duman RS. Regulation of BDNF and trkB mRNA in rat brain by chronic electroconvulsive seizure and antidepressant drug treatments. J Neurosci. 1995;15:7539–47.

  78. 78.

    Thome J, Sakai N, Shin K, Steffen C, Zhang YJ, Impey S, Storm D, Duman RS. cAMP response element-mediated gene transcription is upregulated by chronic antidepressant treatment. J Neurosci. 2000;20:4030–6.

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Acknowledgements

H.S. designed and conducted experiments, analyzed and wrote the paper. N.W., J.S. conducted the experiments in Fig. 3. ACY-738 compound was a gift from Matthew Jarpe, Acetylon Pharmaceutics. Authors would like to thank Prof. Mark Brodie for advice with statistical analysis.

Funding:

VA Merit award-BX001149 (M.M.R.); NIH RO1AT009169 (M.M.R.); NIH P50AA022538 and NIH T32 067631.HS was supported by AHA 16POST27770113

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  1. Department of Physiology and Biophysics, University of Illinois at Chicago, Chicago, IL, 60612, USA

    • Harinder Singh
    • , Nathan Wray
    • , Jeffrey M. Schappi
    •  & Mark M. Rasenick
  2. Department of Psychiatry, University of Illinois at Chicago, Chicago, IL, 60612, USA

    • Mark M. Rasenick
  3. Jesse Brown VAMC, Chicago, IL, 60612, USA

    • Mark M. Rasenick

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Conflict of interest

M.M.R. has received research support from Eli Lilly and Lundbeck, Inc. and is consultant to Otsuka Pharmaceuticals. He also has ownership in Pax Neuroscience. The remaining authors have nothing to disclose. H.S., N.W. and J.S. declare no conflicts of interest.

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Correspondence to Mark M. Rasenick.

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https://doi.org/10.1038/s41386-018-0016-x