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High Ethanol and Acetaldehyde Inhibit Glutamatergic Transmission in the Hippocampus of Aldh2-Knockout and C57BL/6N Mice: an In Vivo and Ex Vivo Analysis


We aimed to investigate whether ethanol (EtOH) and acetaldehyde (AcH) can affect glutamate and its receptors GluN1 and GluA1 in the hippocampus of Aldh2-knockout (Aldh2-KO) and C57BL/6N (wild-type (WT)) mice. To do this, we first examined the effect of local administration of EtOH (100 mM, 200 mM, and 500 mM) and AcH (100 μM, 200 μM, and 500 μM) on extracellular glutamate levels in freely moving mice. Retrodialysis of 200 mM and 500 mM EtOH into the hippocampus of WT and Aldh2-KO mice produced significant decreases in extracellular glutamate levels (p < 0.05). A dose of 500 mM EtOH induced a greater decrease in Aldh2-KO mice (p < 0.05) than in WT mice, indicating the action of AcH. Similarly, perfusion of 200 μM and 500 μM AcH decreased glutamate in Aldh2-KO mice (p < 0.05), but this decrease was not seen in WT mice at any AcH dose. Second, we tested whether the EtOH- and AcH-induced decrease in glutamate was associated with decreases in GluN1 and GluA1 expression, as measured by real-time PCR and Western blot. We found a significant decrease in GluN1 (p < 0.05) and GluA1 (p < 0.05) subunits after a high dose of EtOH (4.0 g/kg) and AcH (200 mg/kg) in WT mice. However, a 2.0 g/kg dose of EtOH did not produce a consistent decrease in GluN1 or GluA1 between messenger RNA and protein. In Aldh2-KO mice, all three doses of EtOH (1.0 g/kg, 2.0 g/kg, and 4.0 g/kg) and AcH (50 mg/kg, 100 mg/kg, and 200 mg/kg) decreased GluN1 expression (p < 0.05), while moderate-to-high doses of EtOH (2.0 g/kg and 4.0 g/kg) and AcH (100 mg/kg and 200 mg/kg) decreased GluA1 expression (p < 0.05). Together, these in vivo and ex vivo data suggest that EtOH and AcH decrease extracellular glutamate in the hippocampus of mice with a concomitant decrease in GluN1 and GluA1 subunits, but these effects require relatively high concentrations and may, therefore, explain the consequences of EtOH intoxication.

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  1. Al-Rejaie S, Dar MS (2006) Behavioral interaction between nicotine and ethanol: possible modulation by mouse cerebellar glutamate. Alcohol Clin Exp Res 30:1223–1233

  2. Ariwodola OJ, Weiner JL (2004) Ethanol potentiation of GABAergic synaptic transmission may be self-limiting: role of presynaptic GABA(B) receptors. J Neurosci 24:10679–10686

  3. Bäckström P, Hyytiä P (2004) Ionotropic glutamate receptor antagonists modulate cue-induced reinstatement of ethanol-seeking behavior. Alcohol Clin Exp Res 28:558–565

  4. Carboni S, Isola R, Gessa GL, Rossetti ZL (1993) Ethanol prevents the glutamate release induced by N-methyl-D-aspartate in the rat striatum. Neurosci Lett 152:133–136

  5. Cha YM, Li Q, Wilson WA, Swartzwelder HS (2006) Sedative and GABAergic effects of ethanol on male and female rats. Alcohol Clin Exp Res 30:113–118

  6. Choi DW (1988) Glutamate neurotoxicity and diseases of the nervous system. Neuron 1:623–634

  7. Chu B, Anantharam V, Treistman SN (1995) Ethanol inhibition of recombinant heteromeric NMDA channels in the presence and absence of modulators. J Neurochem 65:140–148

  8. Cortese BM, Phan KL (2005) The role of glutamate in anxiety and related disorders. CNS Spectr 10:820–830

  9. Dildy JE, Leslie SW (1989) Ethanol inhibits NMDA-induced increases in free intracellular Ca2+ in dissociated brain cells. Brain Res 499:383–387

  10. Ding ZM, Engleman EA, Rodd ZA, McBride WJ (2012) Ethanol increases glutamate neurotransmission in the posterior ventral tegmental area of female Wistar rats. Alcohol Clin Exp Res 36:633–640

  11. Doble A (1999) The role of excitotoxicity in neurodegenerative disease: implications for therapy. Pharmacol Ther 81:163–221

  12. Ericson M, Molander A, Löf E, Engel JA, Söderpalm B (2003) Ethanol elevates accumbal dopamine levels via indirect activation of ventral tegmental nicotinic acetylcholine receptors. Eur J Pharmacol 467:85–93

  13. Faingold CL, N’Gouemo P, Riaz A (1998) Ethanol and neurotransmitter interactions—from molecular to integrative effects. Prog Neurobiol 55:509–535

  14. Fischer W, Franke H, Illes P (2003) Effects of acute ethanol on the Ca2+ response to AMPA in cultured rat cortical GABAergic nonpyramidal neurons. Alcohol Alcohol 38(5):394–399

  15. Grant KA, Lovinger DM (1995) Cellular and behavioral neurobiology of alcohol: receptor-mediated neuronal processes. Clin Neurosci 3(3):155–164

  16. Harris RA (1999) Ethanol actions on multiple ion channels: which are important? Alcohol Clin Exp Res 23:1563–1570

  17. Harrison NL, Skelly MJ, Grosserode EK, Lowes DC, Zeric T, Phister S, Salling MC (2017) Effects of acute alcohol on excitability in the CNS. Neuropharmacology 122:36–45

  18. Hicklin TR, Wu PH, Radcliffe RA, Freund RK, Goebel-Goody SM, Correa PR, Proctor WR, Lombroso PJ, Browning MD (2011) Alcohol inhibition of the NMDA receptor function, long-term potentiation, and fear learning requires striatal-enriched protein tyrosine phosphatase. PNAS 108:6650–6655

  19. Hinoi E, Takarada T, Ueshima T, Tsuchihashi Y, Yoneda Y (2004) Glutamate signaling in peripheral tissues. Eur J Biochem 271:1–13

  20. Hoffman PL, Rabe CS, Moses F, Tabakoff B (1989) N-methyl-D-aspartate receptors and ethanol: inhibition of calcium flux and cyclic GMP production. J Neurochem 52:61937–61940

  21. Izumi Y, Nagashima K, Murayama K, Zorumski CF (2005) Acute effects of ethanol on hippocampal long-term potentiation and long-term depression are mediated by different mechanisms. Neuroscience 136(2):509–517

  22. Jamal M, Ameno K, Uekita I, Kumihashi M, Wang W, Ijiri I (2007) Catalase mediates acetaldehyde formation in the striatum of free-moving rats. Neurotoxicology 28:1245–1248

  23. Jamal M, Ameno K, Miki T, Wang W, Kumihashi M, Isse T, Kawamoto T, Kitagawa K, Nakayama K, Ijiri I, Kinoshita H (2009) Cholinergic alterations following alcohol exposure in the frontal cortex of Aldh2-deficient mice models. Brain Res 1295:37–43

  24. Jamal M, Ameno K, Ruby M, Miki T, Tanaka N, Nakamura Y, Kinoshita H (2013) Ethanol- and acetaldehyde-induced cholinergic imbalance in the hippocampus of Aldh2-knockout mice does not affect nerve growth factor or brain-derived neurotrophic factor. Brain Res 1539:41–47

  25. Jamal M, Ameno K, Miki T, Tanaka N, Ito A, Ono J, Takakura A, Kumihashi M, Kinoshita H (2016a) Ethanol and acetaldehyde differentially alter extracellular dopamine and serotonin in Aldh2-knockout mouse dorsal striatum: a reverse microdialysis study. Neurotoxicology 52:204–209

  26. Jamal M, Ameno K, Tanaka N, Ito A, Takakura A, Kumihashi M, Kinoshita H (2016b) Ethanol and acetaldehyde after intraperitoneal administration to Aldh2-knockout mice-reflection in blood and brain levels. Neurochem Res 41:1029–1034

  27. Jin C, Woodward JJ (2006) Effects of 8 different NR1 splice variants on the ethanol inhibition of recombinant NMDA receptors. Alcohol Clin Exp Res 30:673–679

  28. Kanai Y, Hediger MA (2003) The glutamate and neutral amino acid transporter family: physiological and pharmacological implications. Eur J Pharmacol 479:237–247 Review

  29. Khakpai F, Zarrindast MR, Nasehi M, Haeri-Rohani A, Eidi A (2013) The role of glutamatergic pathway between septum and hippocampus in the memory formation. EXCLI J 12:41–51

  30. Kirihara Y, Takechi M, Kurosaki K, Kobayashi Y, Kurosawa T (2013) Anesthetic effects of a mixture of medetomidine, midazolam and butorphanol in two strains of mice. Exp Anim 62:173–180

  31. Kitagawa K, Kawamoto T, Kunugita N, Tsukiyama T, Okamoto K, Yoshida A, Nakayama K, Nakayama K (2000) Aldehyde dehydrogenase (ALDH) 2 associates with oxidation of methoxyacetaldehyde; in vitro analysis with liver subcellular fraction derived from human and Aldh2 gene targeting mouse. FEBS Lett 476:306–311

  32. Krystal JH, Petrakis IL, Mason G, Trevisan L, D’Souza DC (2003) N-methyl-D-aspartate glutamate receptors and alcoholism: reward, dependence, treatment, and vulnerability. Pharmacol Ther 99:79–94

  33. Lau CG, Takeuchi K, Rodenas-Ruano A, Takayasu Y, Murphy J, Bennett MV, Zukin RS (2009) Regulation of NMDA receptor Ca2+ signalling and synaptic plasticity. Biochem Soc Trans 37(Pt 6):1369–1374

  34. Lima-Landman MT, Albuquerque EX (1989) Ethanol potentiates and blocks NMDA-activated single-channel currents in rat hippocampal pyramidal cells. FEBS Lett 247(1):61–67

  35. Lof E, Chau PP, Stomberg R, Soderpalm B (2007) Ethanol-induced dopamine elevation in the rat—modulatory effects by subchronic treatment with nicotinic drugs. Eur J Pharmacol 555:139–147

  36. Lovinger DM (1993) High ethanol sensitivity of recombinant AMPA-type glutamate receptors expressed in mammalian cells. Neurosci Lett 159:83–87

  37. Lovinger DM, Roberto M (2013) Synaptic effects induced by alcohol. Curr Top Behav Neurosci 13:31–86

  38. Lovinger DM, White G, Weight FF (1989) Ethanol inhibits NMDA-activated ion current in hippocampal neurons. Science 243:1721–1724

  39. Lovinger DM, White G, Weight FF (1990) NMDA receptor-mediated synaptic excitation selectively inhibited by ethanol in hippocampal slice from adult rat. J Neurosci 10(4):1372–1379

  40. Malenka RC, Nicoll RA (1999) Long-term potentiation—a decade of progress? Science 285:1870–1874

  41. Meldrum BS (2000) Glutamate as a neurotransmitter in the brain: review of physiology and pathology. J Nutr 130(4S Suppl):1007S–1015S Review

  42. Miladinovic T, Nashed MG, Singh G (2015) Overview of glutamatergic dysregulation in central pathologies. Biomolecules 5:3112–3141

  43. Mizoi Y, Tatsuno Y, Adachi J, Kogame M, Fukunaga T, Fujiwara S, Hishida S, Ijiri I (1983) Alcohol sensitivity related to polymorphism of alcohol-metabolizing enzymes in Japanese. Pharmacol Biochem Behav 18(Suppl 1):127–133

  44. Moghaddam B, Bolinao ML (1994) Biphasic effect of ethanol on extracellular accumulation of glutamate in the hippocampus and the nucleus accumbens. Neurosci Lett 178:99–102

  45. Möykkynen T, Korpi ER (2012) Acute effects of ethanol on glutamate receptors. Basic Clin Pharmacol Toxicol 111:4–13 Review

  46. Muggironi G, Fois GR, Diana M (2013) Ethanol-derived acetaldehyde: pleasure and pain of alcohol mechanism of action. Front Behav Neurosci 7:87

  47. Nie Z, Madamba SG, Siggins GR (1994) Ethanol inhibits glutamatergic neurotransmission in nucleus accumbens neurons by multiple mechanisms. J Pharmacol Exp Ther 271:1566–1573

  48. Novitskaya YA, Dravolina OA, Zvartau EE, Danysz W, Bespalov AY (2010) Interaction of blockers of ionotropic NMDA receptors and metabotropic glutamate receptors in a working memory test in rats. Neurosci Behav Physiol 40(7):807–811

  49. Paxinos G, Franklin KBJ (2001) The Mouse Brain in Stereotaxic Coordinates, Seconded., Academic Press, San Diego, California

  50. Piepponen TP, Kiianmaa K, Ahtee L (2002) Effects of ethanol on the accumbal output of dopamine, GABA and glutamate in alcohol-tolerant and alcohol-nontolerant rats. Pharmacol Biochem Behav 74:21–30

  51. Quertemont E, Didone V (2006) Role of acetaldehyde in mediating the pharmacological and behavioral effects of alcohol. Alcohol Res Health 29:258–265 Review

  52. Quertemont E, Tambour S (2004) Is ethanol a pro-drug? The role of acetaldehyde in the central effects of ethanol. Trends Pharmacol Sci 25:130–134

  53. Quertemont E, Linotte S, de Witte P (2002) Differential taurine responsiveness to ethanol in high- and low-alcohol sensitive rats: a brain microdialysis study. Eur J Pharmacol 444:143–150

  54. Rawat AK (1974) Brain levels and turnover rates of presumptive neurotransmitters as influenced by administration and withdrawal of ethanol in mice. J Neurochem 22:915–922

  55. Reynolds JD, Brien JF (1994) Effects of acute ethanol exposure on glutamate release in the hippocampus of the fetal and adult guinea pig. Alcohol 11:259–267

  56. Roberto M, Madamba SG, Stouffer DG, Parsons LH, Siggins GR (2004a) Increased GABA release in the central amygdala of ethanol-dependent rats. J Neurosci 45:10159–10166

  57. Roberto M, Schweitzer P, Madamba SG, Stouffer DG, Parsons LH, Siggins GR (2004b) Acute and chronic ethanol alter glutamatergic transmission in rat central amygdala: an in vitro and in vivo analysis. J Neurosci 24(7):1594–1603

  58. Ronald KM, Mirshahi T, Woodward JJ (2001) Ethanol inhibition of N-methyl-D-aspartate receptors is reduced by site-directed mutagenesis of a transmembrane domain phenylalanine residue. J Biol Chem 276:44729–44735

  59. Sanacora G, Treccani G, Popoli M (2012) Towards a glutamate hypothesis of depression: an emerging frontier of neuropsychopharmacology for mood disorders. Neuropharmacology 62:63–77

  60. Sanchis-Segura C, Borchardt T, Vengeliene V, Zghoul T, Bachteler D, Gass P, Sprengel R, Spanagel R (2006) Involvement of the AMPA receptor GluR-C subunit in alcohol-seeking behavior and relapse. J Neurosci 26:1231–1238

  61. Schummers J, Browning MD (2001) Evidence for a role for GABA(A) and NMDA receptors in ethanol inhibition of long-term potentiation. Brain Res 94(1–2):9–14

  62. Shimizu K, Matsubara K, Uezono T, Kimura K, Shiono H (1998) Reduced dorsal hippocampal glutamate release significantly correlates with the spatial memory deficits produced by benzodiazepines and ethanol. Neuroscience 83:701–706

  63. Suzdak PD, Paul SM (1987) Ethanol stimulates GABA receptor-mediated Cl- ion flux in vitro: possible relationship to the anxiolytic and intoxicating actions of alcohol. Psychopharmacol Bull 23:445–451

  64. Tabakoff B, Hoffman PL (1993) Ethanol, sedative hypnotics, and glutamate receptor function in brain and cultured cells. Behav Genet 23:231–236 Review

  65. Tamminga CA, Southcott S, Sacco C, Wagner AD, Ghose S (2012) Glutamate dysfunction in hippocampus: relevance of dentate gyrus and CA3 signaling. Schizophr Bull 38:927–935

  66. Theile JW, Morikawa H, Gonzales RA, Morrisett RA (2008) Ethanol enhances GABAergic transmission onto dopamine neurons in the ventral tegmental area of the rat. Alcohol Clin Exp Res 32:1040–1048

  67. Tiwari V, Veeraiah P, Subramaniam V, Patel AB (2014) Differential effects of ethanol on regional glutamatergic and GABAergic neurotransmitter pathways in mouse brain. J Neurochem 128:628–640

  68. Tsai G, Coyle JT (2002) Glutamatergic mechanisms in schizophrenia. Annu Rev Pharmacol Toxicol 42:165–179 Review

  69. Wall TL, Horn SM, Johnson ML, Smith TL, Carr LG (2000) Hangover symptoms in Asian Americans with variations in the aldehyde dehydrogenase (ALDH2) gene. J Stud Alcohol 61:13–17

  70. Wang R, Reddy PH (2017) Role of glutamate and NMDA receptors in Alzheimer’s disease. J Alzheimers Dis 57:1041–1048

  71. Weiner JL, Dunwiddie TV, Valenzuela CF (1999) Ethanol inhibition of synaptically evoked kainate responses in rat hippocampal CA3 pyramidal neurons. Mol Pharmacol 56(1):85–90

  72. Wirkner K, Eberts C, Poelchen W, Allgaier C, Illes P (2000) Mechanism of inhibition by ethanol of NMDA and AMPA receptor channel functions in cultured rat cortical neurons. Naunyn Schmiedeberg's Arch Pharmacol 362(6):568–576

  73. Xu H, Zhang Y, Ren J (2019) ALDH2 and stroke: a systematic review of the evidence. Adv Exp Med Biol 1193:195–210

  74. Yan QS, Reith ME, Ya SG, Jobe PC (1998) Effect of systemic ethanol on basal and stimulated glutamate releases in the nucleus accumbens of freely moving Sprague-Dawley rats: a microdialysis study. Neurosci Lett 258:29–32

  75. Yang M, Zhang Y, Ren J (2019) ALDH2 polymorphism and ethanol consumption: a genetic-environmental interaction in carcinogenesis. Adv Exp Med Biol 1193:229–236 Review

  76. Zenki KC, Mussulini BH, Rico EP, de Oliveira DL, Rosemberg DB (2014) Effects of ethanol and acetaldehyde in zebrafish brain structures: an in vitro approach on glutamate uptake and on toxicity-related parameters. Toxicol in Vitro 28(5):822–828

  77. Zimatkin SM, Buben AL (2007) Ethanol oxidation in the living brain. Alcohol Alcohol 42(6):529–532

  78. Zorumski CF, Mennerick S, Izumi Y (2014) Acute and chronic effects of ethanol on learning-related synaptic plasticity. Alcohol 48:1–17

  79. Zuo GC, Yang JY, Hao Y, Dong YX, Wu CF (2007) Ethanol and acetaldehyde induce similar changes in extracellular levels of glutamate, taurine and GABA in rat anterior cingulate cortex. Toxicol Lett 169:253–258

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We thank Toyoshi Isse, Toshihiro Kawamoto (University of Occupational and Environmental Health, Fukuoka, Japan), and Kyoko Kitagawa (Hamamatsu University School of Medicine, Japan) for providing the breeding pairs of Aldh2-KO mice.


This work was supported in part by a Grant-in-Aid for Scientific Research (Grant Nos. (c) 22590636 and 20590681) from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Correspondence to Mostofa Jamal.

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Jamal, M., Ito, A., Tanaka, N. et al. High Ethanol and Acetaldehyde Inhibit Glutamatergic Transmission in the Hippocampus of Aldh2-Knockout and C57BL/6N Mice: an In Vivo and Ex Vivo Analysis. Neurotox Res (2020).

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  • Acetaldehyde
  • ALDH2
  • Ethanol
  • Glutamate
  • NMDA and AMPA receptors
  • Hippocampus