Abstract
Prenatal alcohol exposure (PAE) can harm the embryonic development and cause life-long consequences in offspring’s health. To clarify the molecular mechanisms of PAE we have used a mouse model of early alcohol exposure, which is based on maternal ad libitum ingestion of 10% (v/v) ethanol for the first eight days of gestation (GD 0.5–8.5). Owing to the detected postnatal growth-restricted phenotype in the offspring of this mouse model and both prenatal and postnatal growth restriction in alcohol-exposed humans, we focused on imprinted genes Insulin-like growth factor 2 (Igf2), H19, Small Nuclear Ribonucleoprotein Polypeptide N (Snrpn) and Paternally expressed gene 3 (Peg3), which all are known to be involved in embryonic and placental growth and development. We studied the effects of alcohol on DNA methylation level at the Igf2/H19 imprinting control region (ICR), Igf2 differentially methylated region 1, Snrpn ICR and Peg3 ICR in 9.5 embryonic days old (E9.5) embryos and placentas by using MassARRAY EpiTYPER. To determine alcohol-induced alterations globally, we also examined methylation in long interspersed nuclear elements (Line-1) in E9.5 placentas. We did not observe any significant alcohol-induced changes in DNA methylation levels. We explored effects of PAE on gene expression of E9.5 embryos as well as E9.5 and E16.5 placentas by using quantitative PCR. The expression of growth promoter gene Igf2 was decreased in the alcohol-exposed E9.5 and E16.5 placentas. The expression of negative growth controller H19 was significantly increased in the alcohol-exposed E9.5 embryos compared to controls, and conversely, a trend of decreased expression in alcohol-exposed E9.5 and E16.5 placentas were observed. Furthermore, increased Snrpn expression in alcohol-exposed E9.5 embryos was also detected. Our study indicates that albeit no alterations in the DNA methylation levels of studied sequences were detected by EpiTYPER, early PAE can affect the expression of imprinted genes in both developing embryo and placenta.
Highlights
Prenatal alcohol exposure (PAE) can affect the development of embryo and cause a wide variety of birth defects and neuronal disorders
Owing to the relatively moderate alcohol exposure and subtle postnatal growth restriction in this model as well as prenatal growth restriction detected in previous mouse models with early and acute high-level alcohol-exposure [11,12], we cannot exclude the effect of early PAE on the regulation of growth
We examined potential alcohol-induced changes in DNA methylation at the Insulin-like growth factor 2 (Igf2)/H19 imprinting control region (ICR), Igf2 DMR1, Small Nuclear Ribonucleoprotein Polypeptide N (Snrpn) ICR and Paternally expressed gene 3 (Peg3) ICR in E9.5 embryos and placentas
Summary
Prenatal alcohol exposure (PAE) can affect the development of embryo and cause a wide variety of birth defects and neuronal disorders. We have developed a mouse model of early PAE, based on maternal ad libitum ingestion of 10% (v/v) alcohol and took advantage of a mouse strain C57BL/6, which has a strong drinking preference for 10% alcohol [6,7]. The period of this moderate and chronic alcohol exposure is the first eight days of pregnancy (GD 0.5–8.5), from preimplantation to the beginning of neurulation. Owing to the relatively moderate alcohol exposure and subtle postnatal growth restriction in this model as well as prenatal growth restriction detected in previous mouse models with early and acute high-level alcohol-exposure [11,12], we cannot exclude the effect of early PAE on the regulation of growth
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