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.

Inflammatory and oxidative stress-related effects associated with neurotoxicity are maintained after exclusively prenatal trichloroethylene exposure

“…The cerebellum was selected for neurochemical analysis to compare our previous findings using identical doses of TCE with postnatal/early life TCE exposure. In addition, the cerebellum is reportedly sensitive to oxidative stress/redox impairment with toxicant exposure (Giordano et al, 2008; LeDuc et al, 2015). This study provides novel evidence that the effects of prenatal TCE exposure are maintained later in life.…”

Neuronal Transmembrane Chloride Transport Has a Time-Dependent Influence on Survival of Hippocampal Cultures to Oxygen-Glucose Deprivation

“…OGD was induced on DIV 7 as previously described [24,32], by removing CM from the wells and replacing it with a deoxygenated experimental medium without glucose (EM-G), consisting of Neurobasal-A without both glucose and sodium pyruvate (Life Technologies), supplemented with HEPES (Sigma, 10 mM) and l-glutamine (HyClone, 0.5mM). The plates were then immediately placed in a humidified hypoxia chamber (Billups-Rothenberg), which was flushed with 100% N 2 for 10 min, then sealed, and placed in an incubator at 37 • C for 2 h.…”

Echocardiographic and ultrasound evaluation of haemodynamic parameters in hypoxic neonates treated with hypothermia: Study protocol

“…It also seems likely that hypoxic-ischemic encephalopathy (HIE) is secondary to multi-organ dysfunction, primarily cardiopulmonary failure after asphyxia, and is not solely caused by hypoxia ( 16 , 17 ). Because severe perinatal asphyxia often causes severe HIE with long-term outcome, most basic and clinical studies in the field of perinatal asphixia have targeted the central nervous system ( 18 , 19 ).…”