Cellular and Molecular Neurobiology

, Volume 38, Issue 4, pp 955–963 | Cite as

Effects of Acute Alcohol Exposure on Layer 5 Pyramidal Neurons of Juvenile Mice

  • Francesco Ferrini
  • Benjamin Dering
  • Andrea De Giorgio
  • Laura Lossi
  • Alberto Granato
Original Research


Early-onset drinking during childhood or preadolescence is a serious social problem. Yet, most of the basic neurobiological research on the acute effects of ethanol has been carried out on adult or early postnatal animals. We studied the effect of alcohol exposure on the basic electrophysiological properties and cell viability of layer 5 pyramidal neurons from the somatosensory cortex of juvenile (P21–P23) C57BL/6N mice. After bath application of 50 mM ethanol to acute slices of the somatosensory cortex, no adverse effects were detected on cells survival, whereas the input resistance and firing rate of layer 5 neurons were significantly reduced. While the effect on the input resistance was reversible, the depressing effect on cell firing remained stable after 6 min of alcohol exposure. Ethanol application did not result in any significant change of mIPSC frequency, amplitude, and rise time. A slight increase of mIPSC decay time was observed after 6 min of ethanol exposure. The molecular mechanisms leading to these alterations and their significance for the physiology of the cerebral cortex are briefly discussed.


Ethanol Cerebral cortex Electrophysiology Pyramidal neurons Acute slice mIPSC 


Author contributions

FF designed and performed the experiments, analyzed the data, and wrote the manuscript. BD designed and performed the experiments and analyzed the data. ADG performed the experiments and analyzed the data. LL performed the experiments and analyzed the data. AG designed and performed the experiments, analyzed the data, and wrote the manuscript. All the authors have read and approved the manuscript.


  1. Atkinson SE, Williams SR (2009) Postnatal development of dendritic synaptic integration in rat neocortical pyramidal neurons. J Neurophysiol 102:735–751. CrossRefPubMedPubMedCentralGoogle Scholar
  2. Badanich KA, Mulholland PJ, Beckley JT, Trantham-Davidson H, Woodward JJ (2013) Ethanol reduces neuronal excitability of lateral orbitofrontal cortex neurons via a glycine receptor dependent mechanism. Neuropsychopharmacology 38:1176–1188. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Brodie MS, Appel SB (2000) Dopaminergic neurons in the ventral tegmental area of C57BL/6J and DBA/2J mice differ in sensitivity to ethanol excitation. Alcohol Clin Exp Res 24:1120–1124CrossRefPubMedGoogle Scholar
  4. Chen Y, Wu P, Fan X, Chen H, Yang J, Song T, Huang C (2012) Ethanol enhances human hyperpolarization-activated cyclic nucleotide-gated currents. Alcohol Clin Exp Res 36:2036–2046. CrossRefPubMedGoogle Scholar
  5. Cohen EJ, Quarta E, Bravi R, Granato A, Minciacchi D (2017) Neural plasticity and network remodeling: from concepts to pathology. Neuroscience 344:326–345. CrossRefPubMedGoogle Scholar
  6. Criswell HE, Ming Z, Kelm MK, Breese GR (2008) Brain regional differences in the effect of ethanol on GABA release from presynaptic terminals. J Pharmacol Exp Ther 326:596–603. CrossRefPubMedPubMedCentralGoogle Scholar
  7. Day M, Carr DB, Ulrich S, Ilijic E, Tkatch T, Surmeier DJ (2005) Dendritic excitability of mouse frontal cortex pyramidal neurons is shaped by the interaction among HCN, Kir2, and Kleak channels. J Neurosci 25:8776–8787CrossRefPubMedGoogle Scholar
  8. Donovan JE (2009) Estimated blood alcohol concentrations for child and adolescent drinking and their implications for screening instruments. Pediatrics 123:e975–e981. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Donovan JE (2013) The burden of alcohol use: focus on children and preadolescents. Alcohol Res 35:186–192PubMedGoogle Scholar
  10. Dou X, Wilkemeyer MF, Menkari CE, Parnell SE, Sulik KK, Charness ME (2013) Mitogen-activated protein kinase modulates ethanol inhibition of cell adhesion mediated by the L1 neural cell adhesion molecule. Proc Natl Acad Sci USA 110:5683–5688. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Ehrlich D, Pirchl M, Humpel C (2012) Ethanol transiently suppresses choline-acetyltransferase in basal nucleus of Meynert slices. Brain Res 1459:35–42. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Fleming RL, Manis PB, Morrow AL (2009) The effects of acute and chronic ethanol exposure on presynaptic and postsynaptic gamma-aminobutyric acid (GABA) neurotransmission in cultured cortical and hippocampal neurons. Alcohol 43:603–618. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Förstera B, Castro PA, Moraga-Cid G, Aguayo LG (2016) Potentiation of gamma aminobutyric acid receptors (GABAAR) by ethanol: how are inhibitory receptors affected? Front Cell Neurosci 10:114. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Fuenzalida M, Fernandez de Sevilla D, Buño W (2007) Changes of the EPSP waveform regulate the temporal window for spike-timing-dependent plasticity. J Neurosci 27:11940–11948CrossRefPubMedGoogle Scholar
  15. Galindo R, Zamudio PA, Valenzuela CF (2005) Alcohol is a potent stimulant of immature neuronal networks: implications for fetal alcohol spectrum disorder. J Neurochem 94:1500–1511CrossRefPubMedGoogle Scholar
  16. Gilpin NW, Smith AD, Cole M, Weiss F, Koob GF, Richardson HN (2009) Operant behavior and alcohol levels in blood and brain of alcohol-dependent rats. Alcohol Clin Exp Res 33:2113–2123. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Granato A, Palmer LM, De Giorgio A, Tavian D, Larkum ME (2012) Early exposure to alcohol leads to permanent impairment of dendritic excitability in neocortical pyramidal neurons. J Neurosci 32:1377–1382. CrossRefPubMedGoogle Scholar
  18. Han CL, Liao CS, Wu CW, Hwong CL, Lee AR, Yin SJ (1998) Contribution to first-pass metabolism of ethanol and inhibition by ethanol for retinol oxidation in human alcohol dehydrogenase family–implications for etiology of fetal alcohol syndrome and alcohol-related diseases. Eur J Biochem 254:25–31CrossRefPubMedGoogle Scholar
  19. Heit C, Dong H, Chen Y, Shah YM, Thompson DC, Vasiliou V (2015) Transgenic mouse models for alcohol metabolism, toxicity, and cancer. Adv Exp Med Biol 815:375–387. CrossRefPubMedPubMedCentralGoogle Scholar
  20. Hingson RW, Heeren T, Winter MR (2006) Age at drinking onset and alcohol dependence: age at onset, duration, and severity. Arch Pediatr Adolesc Med 160:739–746CrossRefPubMedGoogle Scholar
  21. Huang JJ, Yen CT, Tsai ML, Valenzuela CF, Huang C (2012) Acute ethanol exposure increases firing and induces oscillations in cerebellar Golgi cells of freely moving rats. Alcohol Clin Exp Res 36:2110–2116. CrossRefPubMedPubMedCentralGoogle Scholar
  22. Ikonomidou C, Bittigau P, Ishimaru MJ, Wozniak DF, Koch C, Genz K, Price MT, Stefovska V, Hörster F, Tenkova T, Dikranian K, Olney JW (2000) Ethanol-induced apoptotic neurodegeneration and fetal alcohol syndrome. Science 287:1056–1060CrossRefPubMedGoogle Scholar
  23. Lobo IA, Harris RA (2008) GABA(A) receptors and alcohol. Pharmacol Biochem Behav 90:90–94. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Lotfullina N, Khazipov R (2017) Ethanol and the developing brain: inhibition of neuronal activity and neuroapoptosis. Neuroscientist. PubMedGoogle Scholar
  25. Marszalec W, Aistrup GL, Narahashi T (1998) Ethanol modulation of excitatory and inhibitory synaptic interactions in cultured cortical neurons. Alcohol Clin Exp Res 22:1516–1524CrossRefPubMedGoogle Scholar
  26. McMurray MS, Amodeo LR, Roitman JD (2016) Consequences of adolescent ethanol consumption on risk preference and orbitofrontal cortex encoding of reward. Neuropsychopharmacology 41:1366–1375. CrossRefPubMedGoogle Scholar
  27. Moriguchi S, Zhao X, Marszalec W, Yeh JZ, Narahashi T (2007) Effects of ethanol on excitatory and inhibitory synaptic transmission in rat cortical neurons. Alcohol Clin Exp Res 31:89–99CrossRefPubMedGoogle Scholar
  28. Okamoto T, Harnett MT, Morikawa H (2006) Hyperpolarization-activated cation current (Ih) is an ethanol target in midbrain dopamine neurons of mice. J Neurophysiol 95:619–626CrossRefPubMedGoogle Scholar
  29. Olney JW (2014) Focus on apoptosis to decipher how alcohol and many other drugs disrupt brain development. Front Pediatr 2:81. CrossRefPubMedPubMedCentralGoogle Scholar
  30. Proctor WR, Soldo BL, Allan AM, Dunwiddie TV (1992) Ethanol enhances synaptically evoked GABAA receptor-mediated responses in cerebral cortical neurons in rat brain slices. Brain Res 595:220–227CrossRefPubMedGoogle Scholar
  31. Ramaswamy S, Markram H (2015) Anatomy and physiology of the thick-tufted layer 5 pyramidal neuron. Front Cell Neurosci 9:233. PubMedPubMedCentralGoogle Scholar
  32. Rehm J, Mathers C, Popova S, Thavorncharoensap M, Teerawattananon Y, Patra J (2009) Global burden of disease and injury and economic cost attributable to alcohol use and alcohol-use disorders. Lancet 373:2223–2233. CrossRefPubMedGoogle Scholar
  33. Sanderson JL, Donald Partridge L, Valenzuela CF (2009) Modulation of GABAergic and glutamatergic transmission by ethanol in the developing neocortex: an in vitro test of the excessive inhibition hypothesis of fetal alcohol spectrum disorder. Neuropharmacology 56:541–555. CrossRefPubMedGoogle Scholar
  34. Sessler FM, Hsu FC, Felder TN, Zhai J, Lin RC, Wieland SJ, Kosobud AE (1998) Effects of ethanol on rat somatosensory cortical neurons. Brain Res 804:266–274CrossRefPubMedGoogle Scholar
  35. Siggins GR, Pittman QJ, French ED (1987) Effects of ethanol on CA1 and CA3 pyramidal cells in the hippocampal slice preparation: an intracellular study. Brain Res 414:22–34CrossRefPubMedGoogle Scholar
  36. Smolen TN, Smolen A (1989) Blood and brain ethanol concentrations during absorption and distribution in long-sleep and short-sleep mice. Alcohol 6:33–38CrossRefPubMedGoogle Scholar
  37. Spear LP (2016) Consequences of adolescent use of alcohol and other drugs: studies using rodent models. Neurosci Biobehav Rev 70:228–243. CrossRefPubMedPubMedCentralGoogle Scholar
  38. Tipps ME, Raybuck JD, Lattal KM (2014) Substance abuse, memory, and post-traumatic stress disorder. Neurobiol Learn Mem 112:87–100. CrossRefPubMedGoogle Scholar
  39. Trudell JR, Messing RO, Mayfield J, Harris RA (2014) Alcohol dependence: molecular and behavioral evidence. Trends Pharmacol Sci 35:317–323. CrossRefPubMedPubMedCentralGoogle Scholar
  40. Wei W, Faria LC, Mody I (2004) Low ethanol concentrations selectively augment the tonic inhibition mediated by delta subunit-containing GABAA receptors in hippocampal neurons. J Neurosci 24:8379–8382CrossRefPubMedGoogle Scholar
  41. Weiner JL, Valenzuela CF (2006) Ethanol modulation of GABAergic transmission: the view from the slice. Pharmacol Ther 111:533–554CrossRefPubMedGoogle Scholar
  42. White AM (2003) What happened? Alcohol, memory blackouts, and the brain. Alcohol Res Health 27:186–196PubMedGoogle Scholar
  43. Yan H, Li Q, Fleming R, Madison RD, Wilson WA, Swartzwelder HS (2009) Developmental sensitivity of hippocampal interneurons to ethanol: involvement of the hyperpolarization-activated current, Ih. J Neurophysiol 101:67–83. CrossRefPubMedGoogle Scholar
  44. Zorumski CF, Mennerick S, Izumi Y (2014) Acute and chronic effects of ethanol on learning-related synaptic plasticity. Alcohol 48:1–17. CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of Veterinary SciencesUniversity of TurinGrugliascoItaly
  2. 2.Faculty of Natural SciencesUniversity of StirlingStirlingUK
  3. 3.Faculty of PsychologyUniversity eCampusNovedrateItaly
  4. 4.Department of PsychologyCatholic UniversityMilanItaly

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