The Cerebellum

, Volume 17, Issue 3, pp 326–335 | Cite as

Nicotinamide Inhibits Ethanol-Induced Caspase-3 and PARP-1 Over-activation and Subsequent Neurodegeneration in the Developing Mouse Cerebellum

  • Alessandro IeraciEmail author
  • Daniel G. Herrera
Original Paper


Fetal alcohol spectrum disorder (FASD) is the principal preventable cause of mental retardation in the western countries resulting from alcohol exposure during pregnancy. Ethanol-induced massive neuronal cell death occurs mainly in immature neurons during the brain growth spurt period. The cerebellum is one of the brain areas that are most sensitive to ethanol neurotoxicity. Currently, there is no effective treatment that targets the causes of these disorders and efficient treatments to counteract or reverse FASD are desirable. In this study, we investigated the effects of nicotinamide on ethanol-induced neuronal cell death in the developing cerebellum. Subcutaneous administration of ethanol in postnatal 4-day-old mice induced an over-activation of caspase-3 and PARP-1 followed by a massive neurodegeneration in the developing cerebellum. Interestingly, treatment with nicotinamide, immediately or 2 h after ethanol exposure, diminished caspase-3 and PARP-1 over-activation and reduced ethanol-induced neurodegeneration. Conversely, treatment with 3-aminobenzadine, a specific PARP-1 inhibitor, was able to completely block PARP-1 activation, but not caspase-3 activation or ethanol-induced neurodegeneration in the developing cerebellum. Our results showed that nicotinamide reduces ethanol-induced neuronal cell death and inhibits both caspase-3 and PARP-1 alcohol-induced activation in the developing cerebellum, suggesting that nicotinamide might be a promising and safe neuroprotective agent for treating FASD and other neurodegenerative disorders in the developing brain that shares similar cell death pathways.


Fetal alcohol syndrome Apoptosis Caspase-3 Poly(ADP-ribose) polymerase Developing brain Nicotinamide 



Fetal alcohol spectrum disorder


Fetal alcohol syndrome




Poly (ADP-ribose) polymerase




Polymers of ADP-ribose


Nicotinamide adenine dinucleotide


Nicotinamide adenine dinucleotide phosphate


Postnatal days


Funding Information

A.I. was supported by the De Witt-Reader’s Digest Fellowship, and D.G.H. was supported by grants from the National Alliance for Research on Schizophrenia and Depression and the Reader’s Digest Foundation.

Compliance with Ethical Standards

All animal procedures were approved by the Institutional Animal Care and Use Committees of Weill Cornell Medical College and were conducted in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. 1.
    May PA, Gossage JP, Kalberg WO, Robinson LK, Buckley D, Manning M, et al. Prevalence and epidemiologic characteristics of FASD from various research methods with an emphasis on recent in-school studies. Dev Disabil Res Rev. 2009;15(3):176–92. Scholar
  2. 2.
    Burd L, Klug MG, Martsolf JT, Kerbeshian J. Fetal alcohol syndrome: neuropsychiatric phenomics. Neurotoxicol Teratol. 2003;25(6):697–705. Scholar
  3. 3.
    Bonthius DJ, West JR. Alcohol-induced neuronal loss in developing rats: increased brain damage with binge exposure. Alcohol Clin Exp Res. 1990;14(1):107–18. Scholar
  4. 4.
    Holmes GL. Morphological and physiological maturation of the brain in the neonate and young child. J Clin Neurophysiol. 1986;3(3):209–38. Scholar
  5. 5.
    Ieraci A, Herrera DG. Nicotinamide protects against ethanol-induced apoptotic neurodegeneration in the developing mouse brain. PLoS Med. 2006;3(4):e101. Scholar
  6. 6.
    Ieraci A, Herrera DG. Single alcohol exposure in early life damages hippocampal stem/progenitor cells and reduces adult neurogenesis. Neurobiol Dis. 2007;26(3):597–605. Scholar
  7. 7.
    Ikonomidou C, Bittigau P, Ishimaru MJ, Wozniak DF, Koch C, Genz K, et al. Ethanol-induced apoptotic neurodegeneration and fetal alcohol syndrome. Science. 2000;287(5455):1056–60. Scholar
  8. 8.
    Sowell ER, Jernigan TL, Mattson SN, Riley EP, Sobel DF, Jones KL. Abnormal development of the cerebellar vermis in children prenatally exposed to alcohol: size reduction in lobules I-V. Alcohol Clin Exp Res. 1996;20(1):31–4. Scholar
  9. 9.
    Lubetzky-Vilnai A, Jirikowic TL, McCoy SW. Investigation of the dynamic gait index in children: a pilot study. Pediatr Phys Ther. 2011;23(3):268–73. Scholar
  10. 10.
    Jirikowic TL, McCoy SW, Lubetzky-Vilnai A, Price R, Ciol MA, Kartin D, et al. Sensory control of balance: a comparison of children with fetal alcohol spectrum disorders to children with typical development. J Popul Ther Clin Pharmacol. 2013;20(3):e212–28.PubMedPubMedCentralGoogle Scholar
  11. 11.
    Light KE, Belcher SM, Pierce DR. Time course and manner of Purkinje neuron death following a single ethanol exposure on postnatal day 4 in the developing rat. Neuroscience. 2002;114(2):327–37. Scholar
  12. 12.
    Siler-Marsiglio KI, Madorsky I, Pan Q, Paiva M, Neeley AW, Shaw G, et al. Effects of acute ethanol exposure on regulatory mechanisms of Bcl-2-associated apoptosis promoter, bad, in neonatal rat cerebellum: differential effects during vulnerable and resistant developmental periods. Alcohol Clin Exp Res. 2006;30(6):1031–8. Scholar
  13. 13.
    Siler-Marsiglio KI, Paiva M, Madorsky I, Pan Q, Shaw G, Heaton MB. Functional mechanisms of apoptosis-related proteins in neonatal rat cerebellum are differentially influenced by ethanol at postnatal days 4 and 7. J Neurosci Res. 2005;81(5):632–43. Scholar
  14. 14.
    Bearer CF, Wellmann KA, Tang N, He M, Mooney SM. Choline ameliorates deficits in balance caused by acute neonatal ethanol exposure. Cerebellum. 2015;14(4):413–20. Scholar
  15. 15.
    Cebolla AM, Cheron G, Hourez R, Bearzatto B, Dan B, Servais L. Effects of maternal alcohol consumption during breastfeeding on motor and cerebellar Purkinje cells behavior in mice. Neurosci Lett. 2009;455(1):4–7.
  16. 16.
    Klintsova AY, Goodlett CR, Greenough WT. Therapeutic motor training ameliorates cerebellar effects of postnatal binge alcohol. Neurotoxicol Teratol. 2000;22(1):125–32. Scholar
  17. 17.
    Cheng DT, Jacobson SW, Jacobson JL, Molteno CD, Stanton ME, Desmond JE. Eyeblink classical conditioning in alcoholism and fetal alcohol spectrum disorders. Front Psych. 2015;6:155. Scholar
  18. 18.
    Idrus NM, McGough NNH, Spinetta MJ, Thomas JD, Riley EP. The effects of a single memantine treatment on behavioral alterations associated with binge alcohol exposure in neonatal rats. Neurotoxicol Teratol. 2011;33(4):444–50. Scholar
  19. 19.
    Maiese K, Chong ZZ. Nicotinamide: necessary nutrient emerges as a novel cytoprotectant for the brain. Trends Pharmacol Sci. 2003;24(5):228–32. Scholar
  20. 20.
    Hathorn T, Snyder-Keller A, Messer A. Nicotinamide improves motor deficits and upregulates PGC-1alpha and BDNF gene expression in a mouse model of Huntington’s disease. Neurobiol Dis. 2011;41(1):43–50. Scholar
  21. 21.
    Green KN, Steffan JS, Martinez-Coria H, Sun X, Schreiber SS, Thompson LM, et al. Nicotinamide restores cognition in Alzheimer’s disease transgenic mice via a mechanism involving sirtuin inhibition and selective reduction of Thr231-phosphotau. J Neurosci. 2008;28(45):11500–10. Scholar
  22. 22.
    Yang J, Klaidman LK, Chang ML, Kem S, Sugawara T, Chan P, et al. Nicotinamide therapy protects against both necrosis and apoptosis in a stroke model. Pharmacol Biochem Behav. 2002;73(4):901–10. Scholar
  23. 23.
    Maiese K, Chong ZZ, Hou J, Shang YC. The vitamin nicotinamide: translating nutrition into clinical care. Molecules. 2009;14(9):3446–85. Scholar
  24. 24.
    Alano CC, Garnier P, Ying W, Higashi Y, Kauppinen TM, Swanson RA. NAD+ depletion is necessary and sufficient for poly(ADP-ribose) polymerase-1-mediated neuronal death. J Neurosci. 2010;30(8):2967–78. Scholar
  25. 25.
    Andrabi SA, Dawson TM, Dawson VL. Mitochondrial and nuclear cross talk in cell death: parthanatos. Ann N Y Acad Sci. 2008;1147(1):233–41. Scholar
  26. 26.
    Diaz-Hernandez JI, Moncada S, Bolanos JP, Almeida A. Poly(ADP-ribose) polymerase-1 protects neurons against apoptosis induced by oxidative stress. Cell Death Differ. 2007;14(6):1211–21. Scholar
  27. 27.
    Fatokun AA, Dawson VL, Dawson TM. Parthanatos: mitochondrial-linked mechanisms and therapeutic opportunities. Br J Pharmacol. 2014;171(8):2000–16. Scholar
  28. 28.
    Kauppinen TM, Swanson RA. The role of poly(ADP-ribose) polymerase-1 in CNS disease. Neuroscience. 2007;145(4):1267–72. Scholar
  29. 29.
    Kim MY, Zhang T, Kraus WL. Poly(ADP-ribosyl)ation by PARP-1: “PAR-laying” NAD+ into a nuclear signal. Genes Dev. 2005;19(17):1951–67. Scholar
  30. 30.
    Martire S, Mosca L, D’Erme M. PARP-1 involvement in neurodegeneration: a focus on Alzheimer’s and Parkinson’s diseases. Mech Ageing Dev. 2015;146–148:53–64.
  31. 31.
    Dikranian K, Qin YQ, Labruyere J, Nemmers B, Olney JW. Ethanol-induced neuroapoptosis in the developing rodent cerebellum and related brain stem structures. Brain Res Dev Brain Res. 2005;155(1):1–13. Scholar
  32. 32.
    Luo J. Mechanisms of ethanol-induced death of cerebellar granule cells. Cerebellum. 2012;11(1):145–54. Scholar
  33. 33.
    Cimadamore F, Curchoe CL, Alderson N, Scott F, Salvesen G, Terskikh AV. Nicotinamide rescues human embryonic stem cell-derived neuroectoderm from parthanatic cell death. Stem Cells. 2009;27(8):1772–81. Scholar
  34. 34.
    Yu SW, Wang H, Poitras MF, Coombs C, Bowers WJ, Federoff HJ, et al. Mediation of poly(ADP-ribose) polymerase-1-dependent cell death by apoptosis-inducing factor. Science. 2002;297(5579):259–63. Scholar
  35. 35.
    Outeiro TF, Grammatopoulos TN, Altmann S, Amore A, Standaert DG, Hyman BT, et al. Pharmacological inhibition of PARP-1 reduces alpha-synuclein- and MPP+-induced cytotoxicity in Parkinson’s disease in vitro models. Biochem Biophys Res Commun. 2007;357(3):596–602. Scholar
  36. 36.
    Yokoyama H, Kuroiwa H, Tsukada T, Uchida H, Kato H, Araki T. Poly(ADP-ribose)polymerase inhibitor can attenuate the neuronal death after 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced neurotoxicity in mice. J Neurosci Res. 2010;88(7):1522–36. Scholar
  37. 37.
    Bogan KL, Brenner C. Nicotinic acid, nicotinamide, and nicotinamide riboside: a molecular evaluation of NAD+ precursor vitamins in human nutrition. Annu Rev Nutr. 2008;28(1):115–30. Scholar
  38. 38.
    Goodlett CR, Horn KH, Zhou FC. Alcohol teratogenesis: mechanisms of damage and strategies for intervention. Exp Biol Med. 2005;230(6):394–406. Scholar
  39. 39.
    Guerri C, Bazinet A, Riley EP. Foetal alcohol spectrum disorders and alterations in brain and behaviour. Alcohol Alcohol. 2009;44(2):108–14. Scholar
  40. 40.
    Olney JW, Young C, Wozniak DF, Jevtovic-Todorovic V, Ikonomidou C. Do pediatric drugs cause developing neurons to commit suicide? Trends Pharmacol Sci. 2004;25(3):135–9. Scholar
  41. 41.
    Jevtovic-Todorovic V, Hartman RE, Izumi Y, Benshoff ND, Dikranian K, Zorumski CF, et al. Early exposure to common anesthetic agents causes widespread neurodegeneration in the developing rat brain and persistent learning deficits. J Neurosci. 2003;23(3):876–82.CrossRefPubMedGoogle Scholar
  42. 42.
    Olney JW. Focus on apoptosis to decipher how alcohol and many other drugs disrupt brain development. Front Pediatr. 2014;2:81. Scholar
  43. 43.
    Ullah N, Lee HY, Naseer MI, Ullah I, Suh JW, Kim MO. Nicotinamide inhibits alkylating agent-induced apoptotic neurodegeneration in the developing rat brain. PLoS One. 2011;6(12):e27093. Scholar
  44. 44.
    Hanners NW, Eitson JL, Usui N, Richardson RB, Wexler EM, Konopka G, et al. Western Zika virus in human fetal neural progenitors persists long term with partial cytopathic and limited immunogenic effects. Cell Rep. 2016;15(11):2315–22. Scholar
  45. 45.
    Yuan F, Chen X, Liu J, Feng W, Wu X, Chen S. Up-regulation of Siah1 by ethanol triggers apoptosis in neural crest cells through p38 MAPK-mediated activation of p53 signaling pathway. Arch Toxicol. 2017;91(2):775–84. Scholar
  46. 46.
    Feng Y, Paul IA, LeBlanc MH. Nicotinamide reduces hypoxic ischemic brain injury in the newborn rat. Brain Res Bull. 2006;69(2):117–22. Scholar
  47. 47.
    Ullah N, Ullah I, Lee HY, Naseer MI, Seok PM, Ahmed J, et al. Protective function of nicotinamide against ketamine-induced apoptotic neurodegeneration in the infant rat brain. J Mol Neurosci. 2012;47(1):67–75. Scholar
  48. 48.
    Xu M, Lee EM, Wen Z, Cheng Y, Huang WK, Qian X, et al. Identification of small-molecule inhibitors of Zika virus infection and induced neural cell death via a drug repurposing screen. Nat Med. 2016;22(10):1101–7. Scholar
  49. 49.
    Andrabi SA, Kang HC, Haince JF, Lee YI, Zhang J, Chi Z, et al. Iduna protects the brain from glutamate excitotoxicity and stroke by interfering with poly(ADP-ribose) polymer-induced cell death. Nat Med. 2011;17(6):692–9. Scholar
  50. 50.
    Cherian PP, Schenker S, Henderson GI. Ethanol-mediated DNA damage and PARP-1 apoptotic responses in cultured fetal cortical neurons. Alcohol Clin Exp Res. 2008;32(11):1884–92. Scholar
  51. 51.
    Young C, Roth KA, Klocke BJ, West T, Holtzman DM, Labruyere J, et al. Role of caspase-3 in ethanol-induced developmental neurodegeneration. Neurobiol Dis. 2005;20(2):608–14. Scholar
  52. 52.
    Mukherjee SK, Klaidman LK, Yasharel R, Adams JD. Increased brain NAD prevents neuronal apoptosis in vivo. Eur J Pharmacol. 1997;330(1):27–34. Scholar
  53. 53.
    Walsh SR, Hogg D, Mydlarski PR. Bullous pemphigoid: from bench to bedside. Drugs. 2005;65(7):905–26. Scholar
  54. 54.
    Libri V, Yandim C, Athanasopoulos S, Loyse N, Natisvili T, Law PP, et al. Epigenetic and neurological effects and safety of high-dose nicotinamide in patients with Friedreich’s ataxia: an exploratory, open-label, dose-escalation study. Lancet. 2014;384(9942):504–13. Scholar
  55. 55.
    Olmos PR, Hodgson MI, Maiz A, Manrique M, De Valdes MD, Foncea R, et al. Nicotinamide protected first-phase insulin response (FPIR) and prevented clinical disease in first-degree relatives of type-1 diabetics. Diabetes Res Clin Pract. 2006;71(3):320–33. Scholar
  56. 56.
    Chen AC, Martin AJ, Choy B, Fernandez-Penas P, Dalziell RA, McKenzie CA, et al. A phase 3 randomized trial of nicotinamide for skin-cancer chemoprevention. N Engl J Med. 2015;373(17):1618–26. Scholar
  57. 57.
    Crino A, Schiaffini R, Manfrini S, Mesturino C, Visalli N, Beretta Anguissola G, et al. A randomized trial of nicotinamide and vitamin E in children with recent onset type 1 diabetes (IMDIAB IX). Eur J Endocrinol. 2004;150(5):719–24. Scholar
  58. 58.
    Knip M, Douek IF, Moore WP, Gillmor HA, McLean AE, Bingley PJ, et al. Safety of high-dose nicotinamide: a review. Diabetologia. 2000;43(11):1337–45. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of PsychiatryWeill Medical College of Cornell UniversityNew YorkUSA
  2. 2.Laboratory of Neuropsychopharmacology and Functional Neurogenomics - Dipartimento di Scienze Farmacologiche e Biomolecolari and Center of Excellence on Neurodegenerative DiseasesUniversità di MilanoMilanItaly
  3. 3.Department of Psychiatry Cambridge Health AllianceHarvard Medical School CambridgeCambridgeUSA

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