title: Developmental exposure to ethanol increases the neuronal vulnerability to oxygen-glucose deprivation in cerebellar granule cell cultures.
abstract: Prenatal alcohol exposure is associated with microencephaly, cognitive and behavioral deficits, and growth retardation. Some of the mechanisms of ethanol-induced injury, such as high level oxidative stress and overexpression of pro-apoptotic genes, can increase the sensitivity of fetal neurons towards hypoxic/ischemic stress associated with normal labor. Thus, alcohol-induced sequelae may be the cumulative result of direct ethanol toxicity and increased neuronal vulnerability towards metabolic stressors, including hypoxia. We examined the effects of ethanol exposure on the fetal cerebellar granular neurons' susceptibility to hypoxic/hypoglycemic damage. A chronic ethanol exposure covered the entire prenatal period and 5 days postpartum through breastfeeding, a time interval partially extending into the third-trimester equivalent in humans. After a binge-like alcohol exposure at postnatal day 5, glutamatergic cerebellar granule neurons were cultured and grown for 7 days in vitro, then exposed to a 3-h oxygen-glucose deprivation to mimic a hypoxic/ischemic condition. Cellular viability was monitored by dynamic recording of propidium iodide fluorescence over 20 h reoxygenation. We explored differentially expressed genes on microarray data from a mouse embryonic ethanol-exposure model and validated these by real-time PCR on the present model. In the ethanol-treated cerebellar granule neurons we find an increased expression of genes related to apoptosis (Mapk8 and Bax), but also of genes previously described as neuroprotective (Dhcr24 and Bdnf), which might suggest an actively maintained viability. Our data suggest that neurons exposed to ethanol during development are more vulnerable to in vitro hypoxia/hypoglycemia and have higher intrinsic death susceptibility than unexposed neurons.
authors.affiliation: Division of Physiology and Fundamental Neuroscience, Carol Davila University of Medicine and Pharmacy, 050474 Bucharest, Romania; Molecular Biochemistry, Institute of Biochemistry, Medical Faculty, University of Leipzig, Johannisallee 30, 04103 Leipzig, Germany. Electronic address: diana_leduc@eva.mpg.de.
authors.forename: Diana
authors.initials: D
authors.lastname: Le Duc
authors.affiliation: Division of Physiology and Fundamental Neuroscience, Carol Davila University of Medicine and Pharmacy, 050474 Bucharest, Romania.
authors.forename: Ana
authors.initials: A
authors.lastname: Spataru
authors.affiliation: Division of Physiology and Fundamental Neuroscience, Carol Davila University of Medicine and Pharmacy, 050474 Bucharest, Romania.
authors.forename: Mihai
authors.initials: M
authors.lastname: Ceanga
authors.affiliation: Division of Physiology and Fundamental Neuroscience, Carol Davila University of Medicine and Pharmacy, 050474 Bucharest, Romania.
authors.forename: Leon
authors.initials: L
authors.lastname: Zagrean
authors.affiliation: Molecular Biochemistry, Institute of Biochemistry, Medical Faculty, University of Leipzig, Johannisallee 30, 04103 Leipzig, Germany.
authors.forename: Torsten
authors.initials: T
authors.lastname: Schöneberg
authors.affiliation: Translational Neuroscience, School of Clinical and Experimental Medicine, College of Medical and Dental Sciences, University of Birmingham, Birmingham B15 2TT, United Kingdom.
authors.forename: Emil C
authors.initials: EC
authors.lastname: Toescu
authors.affiliation: Division of Physiology and Fundamental Neuroscience, Carol Davila University of Medicine and Pharmacy, 050474 Bucharest, Romania. Electronic address: azagrean@umf.ro.
authors.forename: Ana-Maria
authors.initials: AM
authors.lastname: Zagrean
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000633:toxicity*; D002531:Cerebellum / Q000473:pathology*; D004622:Embryo, Mammalian; D000431:Ethanol / Q000633:toxicity*; D005260:Female; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000502:physiology; D005947:Glucose / Q000172:deficiency; D000860:Hypoxia / Q000473:pathology; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D020411:Oligonucleotide Array Sequence Analysis; D011247:Pregnancy; D011297:Prenatal Exposure Delayed Effects / Q000139:chemically induced* / Q000473:pathology*; D017208:Rats, Wistar
chemical_list: D002492:Central Nervous System Depressants; D009419:Nerve Tissue Proteins; D000431:Ethanol; D005947:Glucose
keywords: Cerebellum; Differential gene expression; Neuronal vulnerability; Oxygen–glucose deprivation; Prenatal ethanol exposure