Postnatal choline supplementation selectively attenuates hippocampal microRNA alterations associated with developmental alcohol exposure

Abstract

Prenatal alcohol exposure can result in a range of physical, neuropathological, and behavioral alterations, collectively termed fetal alcohol spectrum disorders (FASD). We have shown that supplementation with the nutrient choline reduces the severity of developmental alcohol-associated deficits in hippocampal-dependent behaviors and normalizes some aspects of hippocampal cholinergic development and DNA methylation patterns. Alcohol’s developmental effects may also be mediated, in part, by altering microRNAs (miRNAs) that serve as negative regulators of gene translation. To determine whether choline supplementation alters ethanol’s long-lasting effects on miRNAs, Sprague-Dawley rats were exposed to 5.25 g/kg/day ethanol from postnatal days (PD) 4–9 via intubation; controls received sham intubations. Subjects were treated with choline chloride (100 mg/kg/day) or saline vehicle subcutaneously (s.c.) from PD 4–21. On PD 22, subjects were sacrificed, and RNA isolated from the hippocampus. MiRNA expression was assessed with TaqMan Human MicroRNA Panel Low-Density Arrays. Ethanol significantly increased miRNA expression variance, an effect that was normalized with choline supplementation. Cluster analysis of stably expressed miRNAs that exceeded an ANOVA p<0.05 criterion indicated that for both male and female offspring, control and ethanol-exposed groups were most dissimilar from each other, with choline-supplemented groups in between. MiRNAs that expressed an average 2-fold change due to ethanol exposure were further analyzed to identify which ethanol-sensitive miRNAs were protected by choline supplementation. We found that at a false discovery rate (FDR)-adjusted criterion of p<0.05, miR-200c was induced by ethanol exposure and that choline prevented this effect. Collectively, our data show that choline supplementation can normalize disturbances in miRNA expression following developmental alcohol exposure and can protect specific miRNAs from induction by ethanol. These findings have important implications for the mechanisms by which choline may serve as a potential treatment for FASD.

Persistent Changes in Stress‐Regulatory Genes in Pregnant Women or Children Exposed Prenatally to Alcohol

“…We showed here that choline supplementation in children with an FASD reduced the DNA methylation and increased the expression levels of PER2 and POMC . Our data in children with an FASD are in agreement with the rodent study where choline supplementation during the developmental period normalized PAE‐increased global DNA methylation in the hippocampus and prefrontal cortex (Balaraman et al., ) and Pomc gene methylation in the hypothalamus (Bekdash et al., ). However, it should be mentioned the manner in which choline impacts methylation has varied across studies (Balaraman et al., ; Kovacheva et al., ; Niculescu et al., ).…”

Effect of Choline Supplementation on Neurological, Cognitive, and Behavioral Outcomes in Offspring Arising from Alcohol Exposure During Development: A Quantitative Systematic Review of Clinical and Preclinical Studies

“… Dose calculated based on PD4 average body weight of 13 g reported in Balaraman and colleagues (); each rat injected with 0.1 ml of 18.8 mg/ml; this would be the maximum dose. …”

“…Coles and colleagues () did find that sex was a significant covariate in their generalized linear regression model for analysis of the Bayley Mental Development Index (MDI), but they did not report sex separately in their main data table. However, 5 of the 18 preclinical studies did present data separately for males and females (Balaraman et al., ; Bearer et al., ; Schneider and Thomas, ; Thomas et al., ; Waddell and Mooney, ). Only 1 of these studies (Waddell and Mooney, ) used exposure to alcohol while the placenta was present (gestational day 6 to 20), as opposed to early postnatal exposure during the third‐trimester‐equivalent period.…”

Differential neuroimaging indices in prefrontal white matter in prenatal alcohol‐associated ADHD versus idiopathic ADHD

“…Addition of extracellular choline, on the other hand, protects membranes from drops in PtdCho and other phospholipids (Ulus, Wurtman, Mauron, & Blusztajn, ). In the rat, neonatal and perinatal dietary supplementation with choline affords some apparent protection against the effects of ethanol exposure (Balaraman, Idrus, Miranda, & Thomas, ; Thomas, Abou, & Dominguez, ; Thomas, Biane, O'Bryan, O'Neill, & Dominguez, ; Thomas, Garrison, & O'Neill, ; Thomas, La Fiette, Quinn, & Riley, ; Thomas & Tran, ; Wagner & Hunt, ). Recent results of a long‐anticipated human randomized controlled trial of dietary choline for FASD affirm that the treatment is well tolerated and that it yields at least a limited cognitive benefit (Wozniak et al, , ).…”