Intrauterine Alcohol Exposure Research Articles

Alcohol intake during pregnancy reduces offspring bone epiphyseal growth plate chondrocyte proliferation through transforming growth factor β-1 inhibition in the Sprague Dawley rat humerus.

Intrauterine alcohol exposure delays bone maturation and intensifies osteoporosis and fracture risk. As most studies emphasize the neurological aspects of intrauterine alcohol exposure, there is a lack of research on the implications pertaining to osseous tissue. Previous studies investigated these effects in fetuses, with limited studies on postnatal life. Postnatal studies are crucial since peak bone growth occurs during adolescence. This study aimed at assessing the effects of prenatal alcohol exposure on the humerus proximal and distal growth plate chondrocytes in 3-week-old rats. Sprague Dawley rats (n=9) were assigned to either the ethanol group (n=3), saline (n=3), and untreated (n=3) group and time-mated. Once pregnant, as confirmed by the presence of a copulation plug, the former 2 groups were treated with 0.015 ml/g of 25.2% ethanol and 0.9% saline. The untreated group received no treatment. The left humeri belonging to 6 pups per group were used. Serial sections were cut with a microtome at 5 µm thickness. These sections were stained with haematoxylin and eosin for assessment of normal morphology or immunolabeled with anti-Ki-67 and transforming growth factor β-1 (TGFβ-1) antibody. Prenatal alcohol exposure adversely effected the growth plate sizes and the number of cells in the proliferative zone. Fewer TGFβ-1 immunopositive and proliferative chondrocytes were found using the anti-Ki-67 antibody. This may explain the growth retardation in offspring exposed to gestational alcohol, showing that gestational alcohol exposure inhibits cell proliferation, aiding the diminished stature.

EFFECTS OF CHRONIC MATERNAL ALCOHOLIZATION ON THE CARDIOMYOCYTE CONTRACTILITY IN RAT OFFSPRINGS

The effects of intrauterine alcohol exposure on heart development, specifically the contractile apparatus and myofibril distribution within cardiomyocytes, remain controversial. Aim: This study investigates the postnatal dynamics of myofibril ultrastructure in rat ventricular cardiomyocytes following chronic maternal alcohol exposure. Methods: We analyzed cardiac tissue from offspring of chronically alcoholized rats at various postnatal stages using transmission electron microscopy. Quantitative parameters of cardiomyocyte myofibrils in different ventricular myocardial zones were assessed. Results: Chronic maternal alcohol exposure appeared to inhibit sarcomerogenesis and reduced overall myofibril content in the hearts of newborn offspring. Furthermore, we observed increased myofibril surface area and disrupted spatial orientation in subendocardial left ventricular cardiomyocytes and intramural zones of both ventricles. Even in mature offspring, the myocardial contractile apparatus remained irreversibly damaged, exhibiting reduced myofibril density, increased surface area, and disrupted orientation.

Distal tibial trabecular morphometry in a Sprague Dawley rat model of fetal alcohol syndrome: a micro focus X-ray computed tomography case-control study.

intrauterine alcohol exposure has adverse health effects on the offspring, which may result in fetal Alcohol Spectrum Disorders (FASD). The neurological and craniofacial aspects have been well studied; however, long bones have received limited attention despite the short stature reported in FASD children. time-mated (n=13) pregnant Sprague Dawley dams were assigned to either the ethanol (n=5), saline control (n=5) or untreated group (n=3) which received no treatment. The ethanol and saline control dams were treated with 0.015ml/g of 25.2% ethanol or 0.9% saline, respectively. Treatment was for the first 19 days of gestation. Two pups from each dam were used and terminated at 21 days of age. Paired tibiae were harvested. Each bone was scanned using a Nikon XTH 225L 3D-μCT to investigate trabeculae morphometry. the ethanol group had less bone to total volume (BT/TV), thinnest trabeculae (TbTh) which were less spaced (TbSp) compared to the controls. However, number of trabecular (TbN) remained unaffected in all three groups. Tibial length was similar in all three groups; however, the distal metaphysis volume was smallest in the ethanol group. Logistic regression showed that the distal medullary canal area and trabecular separation were the main parameters affected the most in gestational alcohol. The negative correlation of trabecular thickness and spacing in the ethanol group may be a contributor to bone weakness. gestational alcohol exposure affects bone internal morphology in addition to the bone size. Overall, this study supports the findings of clinical observation of small stature in FAS children.

Tensile strength and trabecular parameter disturbances at 12 weeks postnatally in Sprague Dawley rats exposed to gestational alcohol

Gestational alcohol exposure impairs bone growth and may cause an increase in osteoporosis and fracture risk later in life. The study aimed to investigate how intrauterine alcohol exposure would impede trabecular morphometric parameters and tensile strength in postnatal 12-weekold rats. Time-mated (n=10) pregnant rats were assigned to an ethanol experimental group (n=5) and a saline control group treated with 0.015ml/g of 25.2% ethanol or 0.9% saline for the initial 19 days of pregnancy through oral gavage, respectively. Two pups from each dam were used, and terminated when aged 12-weeks, and ten paired humerus bones and femora obtained per group. The bones were scanned at 20µm resolution. Bone length, the bone volume to total volume ratio (BV/TV), trabecular thickness (TbTh), number (TbN), and spacing (TbSp) were analysed as well as cortical bone architecture. Then, 3-point bending tests were conducted with a universal tensile tester to obtain the maximum force, displacement, and time, as well as the breaking force. Both the humerus and the femur were shorter in the experimental group (ethanol) with a smaller bone fraction area, fewer trabecular that were more widely spaced in both proximal and distal regions although trabecular thickness was similar in both the ethanol and control (saline) groups. Tensile strength revealed group similarities. Therefore, the effects of gestational alcohol exposure were not severe at 12 weeks of age, this may suggest that there is potential skeletal recovery in adult life following intrauterine alcohol exposure. However, gestational alcohol affected trabecular morphometric parameters at 12 weeks of age.

Intrauterine alcohol exposure delays growth and disturbs trabecular morphology in 3-week-old Sprague − Dawley rat femur

Introduction: The consequence of gestational alcohol exposure ranges from stillbirth to miscarriage and fetal alcohol syndrome (FAS). FAS is one of the deleterious causes of congenital disabilities, mental and growth retardation. Several studies suggest that low birth weight and impaired bone growth, as well as a decrease in mineralization in utero, may reduce peak bone mass and increase the risk of fractures and osteoporosis later in life. Therefore, the aim of this study was to determine the effects of intrauterine alcohol exposure on the internal architecture of the femur. Material and Methods: Time-mated (n = 13) pregnant Sprague − Dawley dams were assigned to either the ethanol (n = 5), saline control (n = 5) or untreated control (n = 3) group. The former two groups were treated with 0.015 ml/g of 25.2% ethanol and 0.9% saline for the first 19 days of gestation, respectively. The untreated group received no treatment. Once born, the pups were weaned at 21 days. These rats were then terminated. From each dam, two pups were collected resulting in ethanol (n = 10), saline controls (n = 10), and untreated controls (n = 6). The femora of the pups were dissected and scanned using a 3D-μCT scanner (Nikon XTH 225 L) at 15 μm resolution. Trabecular and cortical parameters were analyzed using Volume Graphics Studio® software following reconstruction. Results: We found altered trabecular parameters in the alcohol exposed group. The diaphyseal cortical and medullary cavity proportions were also affected, particularly in the midshaft. Discussion and Conclusion: These results indicate that gestational alcohol exposure may lower bone structural quality by disturbing the internal morphology of the osseous tissue.

Cortical Neurons and Astrocytes in Intrauterine Ethanol Exposure: The Role of Exposure Time and Postnatal Timeline

Background: The state of cortical neurons and astrocytes are pointers to the health of the brain. These cells are morphologically distorted by alcohol exposure. Intrauterine alcohol exposure remains a challenge with perinatal consequences. The role of exposure time and postnatal timeline on the degree of cortical cell derangement remains a subject of controversy till date. This study therefore examines alcohol exposure and postnatal changes on brain weight, cortical neurons and astrocytes at different developmental periods. Methods: Twenty mature female Wistar rats were time-mated and grouped into 4 groups. Group 1 (control) received distilled water (2 mL/kg), Groups 2, 3 and 4 were administered 2.5 mL/kg of 20% ethanol orally on the 4th, 11th and 18th days of gestation respectively. Rats produced litters and pups' brains were harvested and processed for H&E and Golgi Cox stains at the 3rd and 6th postnatal week. Neuronal and astrocytic densities in the cerebral cortex were evaluated. Results and Conclusion: There was statistically significant increase (p<0.05) in the density of degenerating neurons at the third postnatal week and sixth postnatal weeks in the experimental groups when compared with the control. There was also statistically significant increase (p<0.05) in astrocytic density in groups 2 and 4 at the 3rd postnatal week. There was no statistically significant difference (p>0.05) in the astrocytic densities across groups at the sixth postnatal week. It was concluded that intrauterine alcohol exposure in any of the developmental periods resulted in postnatal neuronal degeneration which persisted till the 6th week. However, increased astrocytic densities is a feature of 1st and 3rd trimester alcohol exposure noted in the 3rd but absent in the 6th postnatal week.

Results of criterional and descriptive assessing dysmorphological disorders associated with prenatal exposure to ethanol in children 7-10 years of age

Diagnosis of disorders that determine the clinical manifestations of fetal alcohol syndrome (FAS) can cause certain difficulties: children with intrauterine alcohol exposure have a fairly wide range of physical, cognitive and behavioral features. The severity of disorders varies, which is determined mainly by the period when mother consumed alcohol (1st, 2nd or 3rd trimester of pregnancy), as well as frequency and amount of alcohol consumed. This article presents results of the second stage of the study among school students 7-10 year of age to identify and assess dysmorphological disorders that have arisen due to prenatal exposure to ethanol. During the first stage of the study, 77 students with delayed physical development (height and weight at the level of the 10th centile and less) were identified with anamnestic data about their mothers’ drinking alcohol during pregnancy. All children were included in the second stage of the study for detailed dysmorphological assessment. Diagnostics was carried out in two directions: criterial, including determining presence or absence of disorders, and if any, determining the severity of facial phenotype of FAS (short palpebral fissure lengths, smooth philtrum, thin upper lip); and descriptive, including determining presence or absence of disorders, and if any, determining severity of other dysmorphological disorders associated with FAS (face shape, features of the eyes, shape of the ears, features of the nostrils and lips, features of the structure of the hands). According to the results of the criteria-based diagnostics, 11 (14.3%) out of 77 children had typical facial anomalies or facial phenotype typical for FAS. 4 children had full correspondence of facial anomalies to severe, and 7 children to moderate. According to results of syndromic diagnosis, all children had from 1 to 6 signs of dysmorphological manifestations of FAS, represented by: upper palmar fold in the form of a «hockey stick» in 42.9%; altered ear configurations in the form of a «railway track» in 42.9%; short palpebral fissure lengths in 41.6%; smooth philtrum in 41.6%; thin upper lip in 41.6%; strabismus in 37.7%; clinodactyly of the 5th finger in 37.7%; small midface in 23.4%.

The association between prenatal alcohol consumption and preschool child stress system disturbance.

Drinking alcohol during pregnancy is considered a risk factor for child development; however, child biomarkers of prenatal alcohol exposure have been rarely studied. We examined whether a meconium alcohol metabolite (ethyl glucuronide, EtG) was associated with child cortisol concentrations at primary school age. For 137 children, prenatal alcohol exposure was operationalized by the meconium biomarker EtG and by maternal self-reports during pregnancy. Two EtG cut-offs (EtG ≥10ng/g and EtG ≥112ng/g) were applied. Cortisol concentrations were measured in saliva and hair samples. Children with EtG ≥10ng/g showed significantly reduced hair cortisol concentrations (HCCs) (p=.050, =0.042). For children with EtG ≥112ng/g, the cortisol awakening response (CAR) was significantly decreased (p=.025, =0.070). These effects were also present in correlational analyses with continuous EtG data, speaking for partly dose-dependent effects. Especially, within the EtG ≥112ng/g group, the basal (CAR: rp =-.642, p=.120) and cumulative (HCC: rp =-.660, p=.107) cortisol parameters were associated with child emotional symptoms at medium effect size. The present study showed both the biological association of intrauterine alcohol exposure with the cortisol stress system, partly dose-dependent, and the functional association with emotional and behavioral symptoms.

Associations of prenatal alcohol exposure and offspring harmful alcohol use: findings from the Raine Study

BackgroundEpidemiological evidence suggests offspring exposed to prenatal alcohol are at increased risk of alcohol use disorders in adulthood. The evidence on the risk of developing harmful alcohol use in adolescence is less clear. MethodsWe used data from the Raine Study, a multi-generational birth cohort study, to examine the association between prenatal alcohol exposure and the risk of harmful alcohol use in offspring at the age of 17 years. Log binomial regression was used to estimate the relative risks (RRs) of harmful alcohol use in offspring exposed to maternal alcohol use in the first (early) and third (late) trimesters of pregnancy. Maternal pre-pregnancy alcohol use was used as a negative control for intrauterine exposure for comparison. ResultsComplete data were available for 1200 mother-offspring pairs. After adjustment for potential confounders, we found increased RRs of harmful alcohol use in offspring born to mothers who consumed four or more standard drinks of alcohol per week during the first trimester [RR 1.45(95% CI: 1.08−1.93)], third trimester [RR 1.34 (95% CI: 1.04–1.72)] and during both trimesters of pregnancy [RR 1.86 (95% CI: 1.16−2.96)]. Maternal pre-pregnancy alcohol use was not associated with an increased risk of harmful alcohol use in offspring [RR 1.15 (95% CI: 0.89−1.48)]. ConclusionObserved associations for maternal prenatal alcohol exposure but not maternal pre-pregnancy alcohol use suggests a biological mechanism for intrauterine alcohol exposure on the risk of harmful alcohol use in the offspring.

Association of Prenatal Alcohol Exposure and Offspring Depression: A Negative Control Analysis of Maternal and Partner Consumption.

BackgroundPrevious research has suggested that intrauterine alcohol exposure is associated with a variety of adverse outcomes in offspring. However, few studies have investigated its association with offspring internalizing disorders in late adolescence.MethodsUsing data from the Avon Longitudinal Study of Parents and Children (ALSPAC), we investigated the associations of maternal drinking in pregnancy with offspring depression at age 18 and 24 (n = 13,480). We also examined partner drinking as a negative control for intrauterine exposure for comparison.ResultsOffspring of mothers that consumed any alcohol at 18 weeks gestation were at increased risk of having a diagnosis of depression (fully adjusted model: OR 1.17, 95% CI 1.02 to 1.34), but there was no clear evidence of association between partners’ alcohol consumption at 18 weeks gestation during pregnancy and increased risk of offspring depression (fully adjusted model: OR 0.87, 95% CI 0.74 to 1.01). Postestimation tests found a positive difference between the association of maternal and partner alcohol use on offspring depression, showing a stronger association for maternal compared with partner alcohol use (OR 1.41, CI 1.07 to 1.84).ConclusionsMaternal drinking in pregnancy was associated with increased risk of offspring depression at age 18. Residual confounding may explain this association, but the negative control comparison of paternal drinking provides some evidence that it may be causal, and this warrants further investigation.

Effects of prenatal alcohol consumption on cognitive development and ADHD-related behaviour in primary-school age: a multilevel study based on meconium ethyl glucuronide.

Alcohol intake during pregnancy is considered to be a risk factor for child development. Child biomarkers of intrauterine alcohol exposure have been rarely studied. We investigated whether a meconium alcohol metabolite (ethyl glucuronide, EtG) was associated with cognitive development, ADHD-related behaviour and neurophysiological markers of attention and executive control of children at primary-school age. Mothers provided self-report on prenatal alcohol consumption during their 3rd trimester. Meconium samples were collected at birth. A total of 44 children with a meconium EtG above the detection limit (≥10ng/g) and 44 nonexposed matched controls were compared. A second threshold (≥154ng/g) was applied to study the dose effects. When children reached primary-school age, mothers rated ADHD-related behaviour, child cognitive development was measured using an IQ test battery, and event-related potentials were recorded during a cued go/nogo task. Children in both EtG-positive groups allocated fewer attentional resources than controls to the go/nogo task (reduced P3 component in go-trials). Children with a meconium EtG above 154ng/g were also found to have an IQ that was six points lower than the other groups. Within the EtG≥154ng/g group, there was a positive correlation between EtG value and ADHD-related behaviour. These significant effects were not observed in relation to the maternal self-report data. Associations between EtG and cognitive deficits, attentional resource capacity and ADHD-related behaviour could be documented with effects that were partially dose-dependent. In addition to maternal self-reports, this biomarker of intrauterine alcohol exposure may be considered as a predictor of child development.

Did you drink alcohol during pregnancy? Inaccuracy and discontinuity of women's self-reports: On the way to establish meconium ethyl glucuronide (EtG) as a biomarker for alcohol consumption during pregnancy.

Consuming alcohol during pregnancy is one of the most verified prenatal risk factors for impaired child development. Information about the amount of alcohol consumed prenatally is needed to anticipate negative effects and to offer timely support. Women's self-reports are not reliable, often influenced by social stigmas and retrospective recall bias, causing biomarkers of intrauterine ethanol exposure to become more and more relevant. The present study compares both women's gestational and retrospective self-reports of prenatal alcohol consumption with levels of ethyl glucuronide (EtG) in meconium. Women (n=180) gave self-reports of prenatal alcohol consumption both during their 3rd trimester (gestational self-report) and when their children were 6-8 years old (retrospective self-report). Child meconium was collected after birth and analyzed for EtG. No individual feedback of children's EtG level was given to the women. All analyses were run separately for two cut-offs: 10ng/g (limit of detection) and 120ng/g (established by Goecke etal., 2014). Mothers of children with EtG values above 10ng/g (n=42) tended to report prenatal alcohol consumption more frequently. There was no trend or significance for the EtG cut-off of 120ng/g (n=26) or for retrospective self-report. When focusing on women who retrospectively reported alcohol consumption during pregnancy, a claim to five or more consumed glasses per month made an EtG over the 10ng/g and the 120ng/g cut-off more probable. Women whose children were over the 10ng/g EtG cut-off were the most inconsistent in their self-report behavior, whereas the consistency in the above 120ng/g EtG group was higher than in any other group. The next step to establish EtG as a biomarker for intrauterine alcohol exposure is to correlate EtG values in meconium with child developmental impairments.

Sleep and melatonin secretion abnormalities in children and adolescents with fetal alcohol spectrum disorders

ObjectivesCaregivers describe significant sleep disturbances in the vast majority of children and adolescents, which is diagnosed as fetal alcohol spectrum disorders (FASD), but objective data on sleep disorders in this population are almost completely lacking. Animal models suggest that intrauterine alcohol exposure may disrupt sleep wake patterns, cause sleep fragmentation, and specifically affect the suprachiasmatic nucleus, thus disrupting melatonin secretion. The objective of this pioneering study was to evaluate sleep and melatonin abnormalities in children with FASD using objective, gold-standard measures. MethodsChildren and adolescents (N = 36, 6–18 years) with FASD participated in clinical assessments by sleep specialists, overnight polysomnography (PSG), and a dim light melatonin onset (DLMO) test in a pediatric sleep laboratory. PSG was analyzed according to standardized scoring guidelines and sleep architecture was compared with normative data. DLMOs were determined and melatonin secretion curves were evaluated qualitatively to classify melatonin profiles. Sleep disorders were evaluated according to international diagnostic criteria. ResultsThere was a high prevalence (58%) of sleep disorders. The most common sleep problems were parasomnias (27.9%) and insomnia (16.8%). The sleep studies showed lower than normal sleep efficiency and high rates of sleep fragmentation. Most participants (79%) had an abnormal melatonin profile. ConclusionsThis study led to the recognition that both sleep and melatonin secretion abnormalities are present in children with FASD. Therefore, to be effective in managing the sleep problems in children with FASD, one needs to consider both the sleep per se and a possible malfunction of the circadian regulation.

Neurological and Psychological Impairments of Children with Fetal Alcohol Spectrum Disorders: Two Case Reports

Background: According to the study “Gesundheit in Deutschland Aktuell”—“Health in Germany up-to-date” (GEDA, Robert-Koch-Institute, 2012), approximately 20% of the pregnant women show moderate and 8% risky alcohol consumption, 12% indicate binge drinking (≥5 drinks per occasion) less than once a month, 4% every month, and 0.1% at least every week. A harmless amount of alcohol for the embryo and fetus cannot be determined based on the scientific state of knowledge. The prevalence of fetal alcohol spectrum disorder (FASD), resulting from the intrauterine alcohol exposure, is estimated to be 1% of all children. Only a fraction of these children really get the diagnosis of FASD.

P-20PERINATES ASSOCIATE ETHANOL'S SENSORY CUES WITH THE DRUG'S DEPRESSIVE RESPIRATORY EFFECTS: LATTER RE-EXPOSURE TO ETHANOL ODOR GENERATES CONDITIONED BREATHING DEPRESSION.

Breathing depressions endanger the neurological wellbeing of human babies. In different species, intrauterine alcohol exposure exerts dramatic effects on fetal breathing patterns. Prior research also shows that perinates smell ethanol in the amniotic fluid following maternal ingestion of the drug and that they associate these cues with different physiological …

Implementation of a Guideline for the Diagnosis of Fetal Alcohol Syndrome

Background: Up to 30% of all pregnant women drink alcohol. The intrauterine alcohol exposure can lead to fetal alcohol syndrome (FAS). FAS has an estimated prevalence of 8 per 1,000 births. Until now, many professional groups are not informed sufficiently about this syndrome and only a fraction of the affected children are diagnosed early in the lifetime. The toxically determined impairments of functions and everyday life can be influenced positively by early and individual support of the patients.

Intrauterine substance exposures and Wechsler Individual Achievement Test-II scores at 11 years of age

The goal of this study was to evaluate whether intrauterine exposure to cocaine, alcohol, tobacco, or marijuana was associated with unique variance in children's academic achievement test scores after controlling for other substance exposures and contextual variables. Academic achievement scores (Wechsler Individual Achievement Test-Second Edition (WIAT-II)) were collected from 119, low-income, urban 11-year-olds enrolled in a prospective longitudinal study of intrauterine cocaine exposure (IUCE). Newborn recruitment eligibility included >36 weeks gestational age, no neonatal intensive care, no intrauterine exposure to illicit drugs except cocaine or marijuana, and no evidence of fetal alcohol syndrome (FAS). Intrauterine substance exposures (cocaine, marijuana, alcohol, tobacco) were documented in the newborn period using biological assays and maternal self-report. Multivariate analyses evaluated the relationship of each intrauterine exposure and each WIAT-II score controlling for contextual variables and the other substance exposures. In secondary analyses, the Child Depression Inventory (CDI) was added to final models. Results indicate that neither IUCE nor intrauterine exposure to marijuana or tobacco was associated with lower WIAT-II scores in multivariable analyses. Intrauterine alcohol exposure predicted significant variance in Math Reasoning and Spelling scores, but inclusion of children's depressive symptoms attenuated the relationship. Lower maternal education and maternal US birth were also associated with lower WIAT-II scores. Our analyses indicate that among low-income, urban 11-year-olds without a diagnosis of FAS, intrauterine alcohol exposure was negatively associated with math and spelling achievement scores after controlling for other intrauterine substance exposures and contextual factors. That relationship in this sample was partially mediated by children's depressive symptoms, but we cannot establish whether these lower achievement scores precede or follow depressive symptoms. Our findings highlight that neurobehavioral studies evaluating a primary potential neuroteratogen should evaluate other associated intrauterine exposures and contextual factors.

Goal‐directed arm movements in children with fetal alcohol syndrome: a kinematic approach

Although many studies have documented deficits in general motor functioning in children with fetal alcohol syndrome (FAS), few have employed detailed measurements to explore the specific nature of such disabilities. This pilot study explores whether three-dimensional (3D) kinematic analysis may generate increased knowledge of the effect of intrauterine alcohol exposure on motor control processes by detecting atypical upper-limb movement pattern specificity in children with FAS relative to typically developing (TD) children. Left and right arm and head movements during a sequential unimanual goal-directed precision task in a sample of children with FAS and in TD children were registered by an optoelectronic tracking system (ProReflex, Qualisys Inc.). Children with FAS demonstrated evidently poorer task performance compared with TD children. Additionally, analyses of arm movement kinematics revealed atypical spatio-temporal organization in the children with FAS. In general, they exhibited longer arm movement trajectories at both the proximal and distal level, faster velocities at the proximal level but slower at the distal level, and more segmented distal movements. Children with FAS also showed atypically augmented and fast head movements during the task performance. Findings indicate neuromotor deficits and developmental delay in goal-directed arm movements because of prenatal alcohol exposure. It is suggested that 3D kinematic analysis is a valid technique for furthering the understanding of motor control processes in children with FAS/fetal alcohol spectrum disorders. A combination with relevant neuroimaging techniques in future studies would enable a more clear-cut interpretation of how atypical movement patterns relate to underlying brain abnormalities.

Candidate Placental Biomarkers for Intrauterine Alcohol Exposure

Fetal alcohol spectrum disorder (FASD) is a leading cause of nongenetic mental retardation and other neurodevelopmental deficits. Earlier diagnosis of FASD would greatly improve prognosis for individuals and families affected by this disorder. Here, we identify candidate placental biomarkers in an animal model of FASD that recapitulates many aspects of human FASD. Pregnant Sprague-Dawley (SD) females were assigned to 1 of 3 diet groups on gestation day 8 (G8): Ethanol (E), Pair-fed (PF) or Control (C). E dams received ethanol-containing liquid diet and PF dams received isocaloric liquid diet in an amount that matched the paired E dam's diet consumption the previous day. Control dams received laboratory chow and water ad libitum. Whole placentae from individual fetuses were collected on gestational day 21 (G21) for analyses. Western blotting and quantitative real-time RT-PCR were used to measure protein and mRNA levels of placental iodothyronine deiodinase III (Dio3), thyroid hormone receptor α1 (TRα1), and glucocorticoid receptor (GR). Placental mRNA levels of insulin-like growth factor 2 (Igf-2), pleckstrin homology-like domain family A member 2 (Phlda2), and cyclin-dependent kinase inhibitor 1C (Cdkn1c) were also measured. Placental protein and mRNA levels from ethanol (E)-consuming dams showed the following changes: increased Dio3, decreased TRα1, and decreased GR compared to both C and PF dams. Placental mRNA levels of intrauterine growth restriction (IUGR) markers Igf-2, Phlda2, and Cdkn1c were altered similarly in PF and E dams. We propose the specific pattern of increased Dio3 and decreased TRα1 and GR protein levels in the placenta as selective biomarker for intrauterine alcohol exposure.

Subtle decreases in DNA methylation and gene expression at the mouse Igf2 locus following prenatal alcohol exposure: effects of a methyl-supplemented diet

C57BL/6J (B6) mice are susceptible to in utero growth retardation and a number of morphological malformations following prenatal alcohol exposure, while DBA/2J (D2) mice are relatively resistant. We have previously shown that genomic imprinting may play a role in differential sensitivity between B6 and D2. The best-characterized mechanism mediating genomic imprinting is differential DNA methylation. In the present study we examined DNA methylation and gene expression, in both embryonic and placental tissue, at the mouse Igf2 locus following in utero ethanol exposure. We also examined the effects of a methyl-supplemented diet on methylation and ethanol teratogenesis. In embryos from susceptible B6 mice, we found small decreases in DNA methylation at four CpG sites in one of the differentially methylated regions of the Igf2 locus; only one of the four sites showed a statistically significant decrease. We observed no significant decreases in methylation in placentae. All Igf2 transcripts showed approximately 1.5-fold decreases following intrauterine alcohol exposure. Placing dams on a methyl-supplemented diet before pregnancy and throughout gestation brought methylation back up to control levels. Methyl supplementation also resulted in lower prenatal mortality, greater prenatal growth, and decreased digit malformations; it dramatically reduced vertebral malformations. Thus, although prenatal alcohol had only small effects on DNA methylation at the Igf2 locus, placing dams on a methyl-supplemented diet partially ameliorated ethanol teratogenesis.