Fetal alcohol syndrome (FAS) is an important cause of mental retardation. Exposure of the developing brain to alcohol can induce the death of neurons, which contributes strongly to learning deficits and neurological problems associated with FAS. Understanding the factors that influence neuronal vulnerability to alcohol induced loss in the developing brain is of considerable significance. We hypothesize that as neurons mature, they acquire a neuroprotective, signaling pathway that protects them against alcohol toxicity. We have discovered a neuroprotective pathway, which is mediated by nitric oxide (NO). This proposal will elucidate the function and molecular mechanisms of the NO-signaling pathway in protecting the developing brain against alcohol-induced neuronal loss. Our experiments will utilize a mouse strain carrying a null mutation for neuronal nitric oxide synthase (nNOS-/-), thus these animals lack NO-mediated neuroprotection.
Specific Aim 1 includes morphological experiments examining the importance of NO-mediated neuroprotection on neuronal survival. Stereological methods will measure alcohol-induced neuronal losses in the hippocampus and cerebellum of mice that express or lack the protective pathway. Alcohol exposure will occur at a variety of ages in order to determine the impact of NO-mediated neuroprotection at different stages of brain development.
Specific Aim 2 utilizes behavioral testing (eyeblink conditioning, Morris Water Maze) to determine whether the enhanced alcohol-induced neuronal losses in nNOS -/- mice are linked to greater functional deficits. Alcohol-induced deficits on these tests will be compared in nNOS / and nNOS+/+ mice and linked with neuronal losses.
Specific Aim 3 includes molecular studies to determine the mechanism by which NO-signaling provides neuroprotection against alcohol toxicity. Experiments will determine whether nNOS is developmentally regulated in vivo and link alcohol vulnerability to expression levels and activity of nNOS. """"""""Rescue"""""""" experiments in which the nNOS gene will be transfected into pathway-deficient neurons in culture derived from nNOS -/- mice will determine whether neuroprotection against alcohol toxicity can be restored. Immunohistochemistry and quantitative RT-PCR will explore the role of cAMP-responsive element binding protein (CREB) as a downstream effector molecule for the NO-signaling and elucidate CREB's role in regulating apoptotic proteins, which may underlie this neuroprotection.

Agency
National Institute of Health (NIH)
Institute
National Institute on Alcohol Abuse and Alcoholism (NIAAA)
Type
Research Project (R01)
Project #
3R01AA011577-08S1
Application #
8023294
Study Section
Neurotoxicology and Alcohol Study Section (NAL)
Program Officer
Hereld, Dale
Project Start
1998-04-01
Project End
2010-12-31
Budget Start
2010-02-15
Budget End
2010-12-31
Support Year
8
Fiscal Year
2010
Total Cost
$34,991
Indirect Cost
Name
University of Iowa
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
062761671
City
Iowa City
State
IA
Country
United States
Zip Code
52242
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Kouzoukas, Dimitrios E; Li, Guiying; Takapoo, Maysaam et al. (2013) Intracellular calcium plays a critical role in the alcohol-mediated death of cerebellar granule neurons. J Neurochem 124:323-35
Karacay, Bahri; Li, Guiying; Pantazis, Nicholas J et al. (2007) Stimulation of the cAMP pathway protects cultured cerebellar granule neurons against alcohol-induced cell death by activating the neuronal nitric oxide synthase (nNOS) gene. Brain Res 1143:34-45
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Bonthius, D J; Pantazis, N J; Karacay, B et al. (2001) Alcohol exposure during the brain growth spurt promotes hippocampal seizures, rapid kindling, and spreading depression. Alcohol Clin Exp Res 25:734-45
Pantazis, N J; Zaheer, A; Dai, D et al. (2000) Transfection of C6 glioma cells with glia maturation factor upregulates brain-derived neurotrophic factor and nerve growth factor: trophic effects and protection against ethanol toxicity in cerebellar granule cells. Brain Res 865:59-76

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