In Vitro Mechanisms of Ethanol Induced Neuronal Death.

Summary

Principal Investigator: Steven J Mennerick
Abstract: Fetal alcohol effects and fetal alcohol syndrome account for a large percentage of metal retardation and behavioral disorders in the United States and impose a tremendous personal and social burden. Understanding how the immature nervous system responds to ethanol is critical to rational intervention strategies. Electrical activity promotes survival of central nervous system (CNS) neurons in vitro and in vivo and prevents natural cell death (NCD) in neurons from many CNS regions. Ethanol depresses CNS electrical activity through interactions with both N-methyl-D-aspartate (NMDA) and gamma-aminobutyric acid (GABA) postsynaptic receptors. Thus it is possible that ethanol enhances developmental NCD. Recent evidence suggests that ethanol exposure is indeed toxic to immature neurons of the forebrain in vivo. The pattern of cell loss is similar to that produced by a combination of glutamate receptor blockade and GABA receptor potentiation. Our evidence suggests that immature hippocampal neurons in vitro are also susceptible to ethanol- induced cell loss, suggesting that susceptibility is intrinsic to neuronal populations affected and that ethanol itself, rather than associated metabolic or nutritional variables, induces the neuronal loss. Hippocampal neurons in culture also die when chronically exposed to GABAmimetics or NMDA receptor blockade. Cell death elicited by all three treatments is prevented by chronically depolarizing neurons. Thus, we have an in vitro model of ethanol-induced neuronal death that will allow us to explore mechanistic questions. We will examine the ultrastructural and biochemical profile of ethanol-induced hippocampal neuronal death in vitro. We will determine whether the interaction of ethanol with NMDA receptors and/or GABA receptors is sufficient to explain the neuronal loss observed in vitro. We will also address whether permanent or acute decreases in calcium signaling are important in cell loss and will determine the time course over which increases in intracellular calcium provide neuroprotection against ethanol-induced cell loss. The proposed experiments should lead to a better fundamental understanding of the mechanisms by which forebrain neurons are susceptible to ethanol-induced death.
Funding Period: 2001-09-30 - 2007-08-31
more information: NIH RePORT

Top Publications

  1. ncbi Astrocytes exert a pro-apoptotic effect on neurons in postnatal hippocampal cultures
    A A Shute
    Departments of Psychiatry and Neurobiology, Washington University School of Medicine, 660 South Euclid Avenue, Campus Box 8134, St Louis, MO 63110, USA
    Neuroscience 131:349-58. 2005
  2. pmc NMDA potentiation by visible light in the presence of a fluorescent neurosteroid analogue
    Lawrence N Eisenman
    Department of Neurology, Washington University School of Medicine, 660 S Euclid Ave, Box 8111, St Louis, MO 63110, USA
    J Physiol 587:2937-47. 2009
  3. pmc Effects on membrane capacitance of steroids with antagonist properties at GABAA receptors
    Steven Mennerick
    Department of Psychiatry, Washington University School of Medicine, St Louis, Missouri 63110, USA
    Biophys J 95:176-85. 2008
  4. pmc Neurosteroid migration to intracellular compartments reduces steroid concentration in the membrane and diminishes GABA-A receptor potentiation
    Ping Li
    Department of Anesthesiology, Washington University School of Medicine, St Louis, MO 63110, USA
    J Physiol 584:789-800. 2007
  5. pmc Antagonism of neurosteroid modulation of native gamma-aminobutyric acid receptors by (3alpha,5alpha)-17-phenylandrost-16-en-3-ol
    Stephen P Kelley
    Department of Psychiatry, University of North Carolina School of Medicine, Chapel Hill, NC, USA
    Eur J Pharmacol 572:94-101. 2007
  6. pmc Mechanisms of neurosteroid interactions with GABA(A) receptors
    Gustav Akk
    Department of Anesthesiology, Washington University School of Medicine, 660 S Euclid Avenue, St Louis, MO 63110, United States
    Pharmacol Ther 116:35-57. 2007
  7. ncbi Anticonvulsant and anesthetic effects of a fluorescent neurosteroid analog activated by visible light
    Lawrence N Eisenman
    Department of Neurology, Washington University School of Medicine, 660 S Euclid Ave, St Louis, Missouri 63110, USA
    Nat Neurosci 10:523-30. 2007
  8. pmc Cyclodextrins sequester neuroactive steroids and differentiate mechanisms that rate limit steroid actions
    H J Shu
    Department of Psychiatry, Washington University School of Medicine, St Louis, MO 63110, USA
    Br J Pharmacol 150:164-75. 2007
  9. pmc A spontaneous tonic chloride conductance in solitary glutamatergic hippocampal neurons
    Lawrence N Eisenman
    Department of Neurology, Washington University School of Medicine, St Louis, MO 63110, USA
    Brain Res 1118:66-74. 2006
  10. pmc Linkage between cellular communications, energy utilization, and proliferation in metastatic neuroendocrine cancers
    Joseph E Ippolito
    Center for Genome Sciences, Washington University, St Louis, MO 63108, USA
    Proc Natl Acad Sci U S A 103:12505-10. 2006

Scientific Experts

  • Lawrence N Eisenman
  • G Akk
  • Steven Mennerick
  • Alana C Conti
  • Krista L Moulder
  • Douglas F Covey
  • Joseph E Ippolito
  • Charles F Zorumski
  • H J Shu
  • Stephen P Kelley
  • Ping Li
  • Kathiresan Krishnan
  • C F Zorumski
  • Xiaoping Jiang
  • John W Olney
  • Jeffrey I Gordon
  • James W Maas
  • A A Shute
  • Julian P Meeks
  • Sriyani Pathirathna
  • Ann Benz
  • Chun Min Zeng
  • C Wang
  • D F Covey
  • Joe Henry Steinbach
  • A Leslie Morrow
  • Todd K O'Buckley
  • C M Zeng
  • Hong Jin Shu
  • Cunde Wang
  • Jamie K Alan
  • Amanda A Taylor
  • Fredrik Backhed
  • Matthew E Merritt
  • Seth T Gammon
  • Jill K Manchester
  • David Piwnica-Worms
  • Xin Jiang
  • K E Isenberg
  • Sherri K Vogt
  • A Benz
  • R J Cormier
  • Louis J Muglia
  • Slobodan M Todorovic
  • Miljen M Jagodic
  • Sanjay Jain
  • Tim West
  • Amanda A Shute
  • Alex S Evers
  • Ricardo A Indacochea
  • Lisa M Muglia
  • Jan R Crowley
  • Jian Xu
  • Vesna Jevtovic-Todorovic
  • Barbara C Brimelow
  • R Reid Townsend
  • Timothy T Tran
  • K L Moulder
  • David M Holtzman
  • Brad D Manion
  • Krista Moulder

Detail Information

Publications19

  1. ncbi Astrocytes exert a pro-apoptotic effect on neurons in postnatal hippocampal cultures
    A A Shute
    Departments of Psychiatry and Neurobiology, Washington University School of Medicine, 660 South Euclid Avenue, Campus Box 8134, St Louis, MO 63110, USA
    Neuroscience 131:349-58. 2005
    ..In sum, our results suggest the surprising result that astrocytes can be negative modulators of neuronal survival during development and when the immature nervous system is challenged with drugs that dampen electrical excitability...
  2. pmc NMDA potentiation by visible light in the presence of a fluorescent neurosteroid analogue
    Lawrence N Eisenman
    Department of Neurology, Washington University School of Medicine, 660 S Euclid Ave, Box 8111, St Louis, MO 63110, USA
    J Physiol 587:2937-47. 2009
    ..Collectively, these data suggest that visible-light potentiation of NMDA receptors by fluorescence excitation shares mechanisms with UV and redox potentiation and may involve singlet oxygen production...
  3. pmc Effects on membrane capacitance of steroids with antagonist properties at GABAA receptors
    Steven Mennerick
    Department of Psychiatry, Washington University School of Medicine, St Louis, Missouri 63110, USA
    Biophys J 95:176-85. 2008
    ..These findings suggest that negatively charged sulfated steroids alter the plasma membrane capacitance without physical movement of the molecule through the electric field...
  4. pmc Neurosteroid migration to intracellular compartments reduces steroid concentration in the membrane and diminishes GABA-A receptor potentiation
    Ping Li
    Department of Anesthesiology, Washington University School of Medicine, St Louis, MO 63110, USA
    J Physiol 584:789-800. 2007
    ..The findings suggest that previous studies overestimate the minimum concentration of steroid capable of potentiating GABA actions at GABA-A receptors. The results have implications for the physiological role of endogenous neurosteroids...
  5. pmc Antagonism of neurosteroid modulation of native gamma-aminobutyric acid receptors by (3alpha,5alpha)-17-phenylandrost-16-en-3-ol
    Stephen P Kelley
    Department of Psychiatry, University of North Carolina School of Medicine, Chapel Hill, NC, USA
    Eur J Pharmacol 572:94-101. 2007
    ..alphaxalone in recombinant alpha1beta2gamma2 receptors. These data provide further evidence of the specificity of 17PA and the heterogeneity of neurosteroid recognition sites on GABAA receptors in the CNS...
  6. pmc Mechanisms of neurosteroid interactions with GABA(A) receptors
    Gustav Akk
    Department of Anesthesiology, Washington University School of Medicine, 660 S Euclid Avenue, St Louis, MO 63110, United States
    Pharmacol Ther 116:35-57. 2007
    ..Using these tools, combined with receptor binding and electrophysiological assays, we have begun to untangle the complexity of steroid actions at this important class of ligand-gated ion channel...
  7. ncbi Anticonvulsant and anesthetic effects of a fluorescent neurosteroid analog activated by visible light
    Lawrence N Eisenman
    Department of Neurology, Washington University School of Medicine, 660 S Euclid Ave, St Louis, Missouri 63110, USA
    Nat Neurosci 10:523-30. 2007
    ..Fluorescent steroids photoactivated by visible light may be useful for modulating GABAA receptor function in a spatiotemporally defined manner...
  8. pmc Cyclodextrins sequester neuroactive steroids and differentiate mechanisms that rate limit steroid actions
    H J Shu
    Department of Psychiatry, Washington University School of Medicine, St Louis, MO 63110, USA
    Br J Pharmacol 150:164-75. 2007
    ....
  9. pmc A spontaneous tonic chloride conductance in solitary glutamatergic hippocampal neurons
    Lawrence N Eisenman
    Department of Neurology, Washington University School of Medicine, St Louis, MO 63110, USA
    Brain Res 1118:66-74. 2006
    ..We conclude that this bicuculline-sensitive conductance needs to be accounted for in studies of GABA tonic currents, lest it be confused with currents associated with GABA overflow...
  10. pmc Linkage between cellular communications, energy utilization, and proliferation in metastatic neuroendocrine cancers
    Joseph E Ippolito
    Center for Genome Sciences, Washington University, St Louis, MO 63108, USA
    Proc Natl Acad Sci U S A 103:12505-10. 2006
    ....
  11. ncbi Physiological activity depresses synaptic function through an effect on vesicle priming
    Krista L Moulder
    Department of Psychiatry, Washington University School of Medicine, St Louis, Missouri 63110, USA
    J Neurosci 26:6618-26. 2006
    ..These results show that glutamatergic neurotransmission persistently adapts to changes in electrical activity over a wide physiological range...
  12. ncbi Synaptic vesicles: turning reluctance into action
    Krista L Moulder
    Department of Psychiatry, Washington University School of Medicine, St Louis, MO 63310, USA
    Neuroscientist 12:11-5. 2006
    ..Recent work from the calyx of Held synapse suggests that reluctance might arise from inactivation of Ca(2+) channels. The authors review this work, along with several other potential mechanisms of reluctance...
  13. ncbi Neurosteroid access to the GABAA receptor
    Gustav Akk
    Department of Anesthesiology, Washington University, St Louis, Missouri 63110, USA
    J Neurosci 25:11605-13. 2005
    ..Thus, neuroactive steroids do not require direct aqueous access to the receptor, and membrane accumulation is required for receptor modulation...
  14. pmc An integrated functional genomics and metabolomics approach for defining poor prognosis in human neuroendocrine cancers
    Joseph E Ippolito
    Center for Genome Sciences and Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St Louis, MO 63110, USA
    Proc Natl Acad Sci U S A 102:9901-6. 2005
    ..Transcriptional, metabolic, and electrophysiologic features of transformed mouse NE cells are also evident in neural progenitor cells...
  15. pmc Action potential fidelity during normal and epileptiform activity in paired soma-axon recordings from rat hippocampus
    Julian P Meeks
    Department of Psychiatry, Washington University School of Medicine, St Louis, MO 63110, USA
    J Physiol 566:425-41. 2005
    ..Such changes in axonal propagation properties could encode information and/or serve as an endogenous brake on seizure propagation...
  16. ncbi Neurosteroid analogues. 10. The effect of methyl group substitution at the C-6 and C-7 positions on the GABA modulatory and anesthetic actions of (3alpha,5alpha)- and (3alpha,5beta)-3-hydroxypregnan-20-one
    Chun Min Zeng
    Department of Molecular Biology and Pharmacology, Washington University School of Medicine, 660 South Euclid Avenue, St Louis, Missouri 63110, USA
    J Med Chem 48:3051-9. 2005
    ....
  17. ncbi New evidence that both T-type calcium channels and GABAA channels are responsible for the potent peripheral analgesic effects of 5alpha-reduced neuroactive steroids
    Sriyani Pathirathna
    Department of Anesthesiology, University of Virginia Health System, Charlottesville, VA, USA
    Pain 114:429-43. 2005
    ..Our results also reveal a role of GABA-gated ion channels in peripheral nociceptive signaling...
  18. ncbi Calcium-stimulated adenylyl cyclases modulate ethanol-induced neurodegeneration in the neonatal brain
    James W Maas
    Department of Pediatrics, Washington University School of Medicine, St Louis, Missouri 63110, USA
    J Neurosci 25:2376-85. 2005
    ..Thus, AC1 and AC8 are critical modulators of neurodegeneration induced by activity blockade in the neonatal brain and represent genetic loci that may potentially modify the severity of fetal alcohol syndrome...
  19. pmc Adenylyl cyclases 1 and 8 initiate a presynaptic homeostatic response to ethanol treatment
    Alana C Conti
    Department of Pediatrics, Washington University in St Louis, St Louis, Missouri, United States of America
    PLoS ONE 4:e5697. 2009
    ..While not direct targets for ethanol, we hypothesize that these cyclases initiate a homeostatic presynaptic response by PKA to reactivate neurons from ethanol-mediated inhibition...