Supplementary Material for: Third Trimester-Equivalent Ethanol Exposure Is Characterized by an Acute Cellular Stress Response and an Ontogenetic Disruption of Genes Critical for Synaptic Establishment and Function in Mice

The developing brain is remarkably sensitive to alcohol exposure,
resulting in the wide range of cognitive and neurobehavioral
characteristics categorized under the term fetal alcohol spectrum
disorders (FASD). The brain is particularly susceptible to alcohol
during synaptogenesis, a process that occurs heavily during the third
trimester and is characterized by the establishment and pruning of
neural circuitry; however, the molecular response of the brain to
ethanol during synaptogenesis has not been documented. To model a
binge-like exposure during the third-trimester neurodevelopmental
equivalent, neonate mice were given a high (5 g/kg over 2 h) dose of
ethanol at postnatal day 7. Acute transcript changes within the brain
were assessed using expression arrays and analyzed for associations with
gene ontology functional categories, canonical pathways, and gene
network interactions. The short-term effect of ethanol was characterized
by an acute stress response and a downregulation of energetically
costly cellular processes. Further, alterations to a number of genes
with roles in synaptic transmission and hormonal signaling, particularly
those associated with the neuroendocrine development and function, were
evident. Ethanol exposure during synaptogenesis was also associated
with altered histone deacetylase and microRNA transcript levels,
suggesting that abnormal epigenetic patterning may maintain some of the
persistent molecular consequences of developmental ethanol exposure. The
results shed insight into the sensitivity of the brain to ethanol
during the third-trimester equivalent and outline how ethanol-induced
alterations to genes associated with neural connectivity may contribute
to FASD phenotypes.