Prenatal alcohol exposure alters expression of neurogenesis-related genes in an ex vivo cell culture model

Abstract

Prenatal alcohol exposure can lead to long-lasting changes in functional and genetic programs of the brain, which may underlie behavioral alterations seen in Fetal Alcohol Spectrum Disorder (FASD). Aberrant fetal programming during gestational alcohol exposure is a possible mechanism by which alcohol imparts teratogenic effects on the brain; however, current methods used to investigate the effects of alcohol on development often rely on either direct application of alcohol in vitro or acute high doses in vivo. In this study, we used our established moderate prenatal alcohol exposure (PAE) model, resulting in maternal blood alcohol content of approximately 20 mM, and subsequent ex vivo cell culture to assess expression of genes related to neurogenesis. Proliferating and differentiating neural progenitor cell culture conditions were established from telencephalic tissue derived from embryonic day (E) 15–17 tissue exposed to alcohol via maternal drinking throughout pregnancy. Gene expression analysis on mRNA derived in vitro was performed using a microarray, and quantitative PCR was conducted for genes to validate the microarray. Student's t tests were performed for statistical comparison of each exposure under each culture condition using a 95% confidence interval. Eleven percent of genes on the array had significantly altered mRNA expression in the prenatal alcohol-exposed neural progenitor culture under proliferating conditions. These include reduced expression of Adora2a, Cxcl1, Dlg4, Hes1, Nptx1, and Vegfa and increased expression of Fgf13, Ndn, and Sox3; bioinformatics analysis indicated that these genes are involved in cell growth and proliferation. Decreased levels of Dnmt1 and Dnmt3a were also found under proliferating conditions. Under differentiating conditions, 7.3% of genes had decreased mRNA expression; these include Cdk5rap3, Gdnf, Hey2, Heyl, Pard6b, and Ptn, which are associated with survival and differentiation as indicated by bioinformatics analysis. This study is the first to use chronic low to moderate PAE, to more accurately reflect maternal alcohol consumption, and subsequent neural progenitor cell culture to demonstrate that PAE throughout gestation alters expression of genes involved in neural development and embryonic neurogenesis.

Prenatal Alcohol Exposure Affects Progenitor Cell Numbers in Olfactory Bulbs and Dentate Gyrus of Vervet Monkeys

“…There are many leads, but few directed or hypothesis-testing studies. Fetal alcohol exposure alters the expression of proteins involved in cell growth, proliferation, survival and differentiation in ex vivo model systems [77,78]. Prenatal ethanol exposure has also been proposed to disrupt the developmental cycle of normal stem cells [79,80].…”

Prenatal arsenic exposure alters REST/NRSF and microRNA regulators of embryonic neural stem cell fate in a sex-dependent manner

“…Neural progenitor cell proliferation and differentiation cultures were generated from telencephalic tissue derived from embryonic day (E) 14 as previously described [58]. Briefly, embryos from control and arsenic-exposed dams were removed at gestational day 14.…”

Modeling Fetal Alcohol Spectrum Disorder: Validating an Ex Vivo Primary Hippocampal Cell Culture System

“…Direct in vitro administration of E can capture the teratogenic toxic effects (Yanni and Lindsley, 2000), but it lacks the ability to recapitulate physiological and indirect consequences of gestational E exposure. This drawback of the in vitro FASD models is resolved in a recent ex vivo study, where E administration occurs in vivo and the evaluation of gene expression occurs ex vivo (Tyler and Allan, 2014). The authors show that moderate prenatal E exposure alters the expression of several genes involved in neurogenesis.…”