Model Of Fetal Alcohol Syndrome Research Articles

Hesperidin Protects Alcohol-Induced Mitochondrial Abnormalities via the Inhibition of Oxidative Stress and MPT Pore Opening in Newborn Male Rats as a Fetal Alcohol Syndrome Model.

Prenatal alcohol exposure causes fetal developmental abnormalities via mitochondrial dysfunction, reactive oxygen species (ROS) formation, and oxidative stress. Therefore, we aimed to investigate the potential of hesperidin as a mitochondrial protective and antioxidative agent in newborn male rats as a model for fetal alcohol syndrome (FAS). Newborn male rats were divided randomly into five groups: a sham group (receiving 27.8 ml/ kg milk solution, orally), an ethanol group (5.25 g/kg in milk solution, orally, 2-10 days after birth), an ethanol + hesperidin group (25 mg/kg/ day orally), an ethanol + hesperidin group (50 mg/kg/day orally), and an ethanol + hesperidin group (100 mg/kg/day orally). Thirty-six days after birth, newborn male rats were sacrificed and brain mitochondria were isolated using differential centrifugation. Mitochondrial toxicity biomarkers of succinate dehydrogenase (SDH) activity, mitochondrial swelling, mitochondrial membrane potential (MMP), and ROS were measured. Offspring neonatally exposed to ethanol showed a significant reduction in SDH activity, mitochondrial swelling, MMP collapse, induction of ROS formation, and lipid peroxidation in isolated mitochondria. Oral administration of hesperidin restored SDH activity, improved MMP collapse and mitochondrial swelling, and reduced ROS formation. This study demonstrates that hesperidin exerts a potent protective effect against alcohol-induced mitochondrial toxicity in the FAS model. Moreover, these findings indicate that hesperidin might be a useful compound for prevention of alcohol-induced fetal developmental abnormalities during pregnancy.

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.

Oligodendrocyte pathology in fetal alcohol spectrum disorders

The pathology of fetal alcohol syndrome and the less severe fetal alcohol spectrum disorders includes brain dysmyelination. Recent studies have shed light on the molecular mechanisms underlying these white matter abnormalities. Rodent models of fetal alcohol syndrome and human studies have shown suppressed oligodendrocyte differentiation and apoptosis of oligodendrocyte precursor cells. Ethanol exposure led to reduced expression of myelin basic protein and delayed myelin basic protein expression in rat and mouse models of fetal alcohol syndrome and in human histopathological specimens. Several studies have reported increased expression of many chemokines in dysmyelinating disorders in central nervous system, including multiple sclerosis and fetal alcohol syndrome. Acute ethanol exposure reduced levels of the neuroprotective insulin-like growth factor-1 in fetal and maternal sheep and in human fetal brain tissues, while ethanol increased the expression of tumor necrosis factor α in mouse and human neurons. White matter lesions have been induced in the developing sheep brain by alcohol exposure in early gestation. Rat fetal alcohol syndrome models have shown reduced axon diameters, with thinner myelin sheaths, as well as reduced numbers of oligodendrocytes, which were also morphologically aberrant oligodendrocytes. Expressions of markers for mature myelination, including myelin basic protein, also were reduced. The accumulating knowledge concerning the mechanisms of ethanol-induced dysmyelination could lead to the development of strategies to prevent dysmyelination in children exposed to ethanol during fetal development. Future studies using fetal oligodendrocyte- and oligodendrocyte precursor cell-derived exosomes isolated from the mother’s blood may identify biomarkers for fetal alcohol syndrome and even implicate epigenetic changes in early development that affect oligodendrocyte precursor cell and oligodendrocyte function in adulthood. By combining various imaging modalities with molecular studies, it may be possible to determine which fetuses are at risk and to intervene therapeutically early in the pregnancy.

Imaging to Detect Abnormalities in the Innervation and Microvasculature of the Developing Heart

The developing heart is a challenge to image because of its small size, rapid morphogenesis, and complex 3‐D structure. Capturing function is even more challenging because the heart begins beating at an early stage. Our group has developed a suite of biophotonic tools to capture structure and function in a quail model of fetal alcohol syndrome, induced by a single administration of ethanol to the fertilized egg to mimic a first trimester binge drinking episode. The quail is a useful model because it develops a 4‐chambered heart, it is economical, and the embryos are more easily accessible than mouse embryos to biophotonic interrogation for longitudinal structure and function analysis. Ethanol exposure reduces survival and increases the prevalence of congenital malformations, including congenital heart defects. We have previously reported on structural heart defects in the outflow tract, great vessels, septation, and in cardiac valves, and we have discovered that certain compounds (e.g. betaine) can alleviate these negative effects.Prenatal ethanol is thought to induce cardiac abnormalities by damaging neural crest cells, which may cause abnormalities in their derivatives. Many studies, including ours, have focused on the consequences of neural crest cell disruption in the formation of abnormalities of the cardiac outflow tract. In our current studies we explore how ethanol exposure may disrupt cardiac innervation, coronary vasculature, and other important derivatives of the neural crest cells.Eggs were injected with an ethanol at gastrulation and incubated until the embryos form four chambered hearts. The heart were dissected and immunofluorescently stained as whole mounts with neuron‐specific tubulin (TUJ1) to label the autonomic innervation. Our preliminary findings demonstrate that ethanol‐treated embryos have fewer innervation branching points and ganglia on the ventral ventricular base than controls. We also found more splitting of the major nerve, the thoracic sympathetic cardiac branch of the vagus that runs along the ventral surface of the pulmonary artery. Filling the microvasculature with SLIME (Scattering Labeled Imaging of Microvascular in Excised tissues) revealed that ethanol exposure results in coronary microvascular misalignment and uneven vessel distribution. In summary, both innervation and coronary microvasculature of the heart are altered by ethanol exposure. These findings may have implications for the cardiac health risks of individuals exposed to ethanol prenatally.Support or Funding InformationNIH R01 HL126747, NIH R01 HL083048, NIH T32 HD060537This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Effects of ethanol and varenicline on female Sprague-Dawley rats in a third trimester model of fetal alcohol syndrome.

Perinatal ethanolexposure disrupts a variety of developmental processes in neurons important for establishing a healthy brain. These ethanol-induced impairments known as fetal alcohol spectrum disorder (FASD) are not fully understood, and currently, there is no effective treatment. Further, growing evidence suggests that adult females are more susceptible to ethanol, with the effects of perinatal ethanol exposure also being sexually divergent. Female models have been historically underutilized in neurophysiological investigations, but here, we used a third-trimester binge-ethanol model of FASD to examine changes to basal forebrain (BF) physiology and behavior in female Sprague-Dawley rats. We also tested varenicline as a potential cholinomimetic therapeutic. Rat pups were gavage-treated with binge-like ethanol, varenicline and ethanol, and varenicline alone. Using patch-clamp electrophysiology in BF slices, we observed that binge-ethanol exposure increased spontaneous post-synaptic current (sPSC) frequency. Varenicline exposure alone also enhanced sPSC frequency. Varenicline plus ethanol co-treatment prevented the sPSC frequency increase. Changes in BF synaptic transmission persisted into adolescence after binge-ethanol treatment. Behaviorally, binge-ethanol treated females displayed increased anxiety (thigmotaxis) and demonstrated learning deficits in the water maze. Varenicline/ethanol co-treatment was effective at reducing these behavioral deficits. In the open field, ethanol-treated rats displayed longer distances traveled and spent less time in the center of the open field box. Co-treated rats displayed less anxiety, demonstrating a possible effect of varenicline on this measure. In conclusion, ethanol-induced changes in both BF synaptic transmission and behavior were reduced by varenicline in female rats, supporting a role for cholinergic therapeutics in FASD treatment.

Skeletal examination in a chick embryo model of fetal alcohol syndrome indicates impaired osteogenesis

In this chapter, we study Riemannian maps from almost Hermitian manifolds to Riemannian manifolds. In section 1, we study holomorphic Riemannian maps as a generalization of holomorphic submersions and obtain a characterization of such maps. In section 2, we investigate anti-invariant Riemannian maps as a generalization of anti-invariant submersion, investigate the geometry of leaves of distributions defined by such maps, and give necessary and sufficient conditions for anti-invariant Riemannian maps to be totally geodesic. We also find necessary and sufficient conditions for the total manifold of anti-invariant Riemannian maps to be an Einstein manifold. In section 3, as a generalization of the semi-invariant submersion, we introduce semi-invariant Riemannian maps, give examples, and obtain the main properties of such maps. In section 4, we introduce generic Riemannian maps from almost Hermitian manifolds to Riemannian manifolds as a generalization of generic submersions and we give examples. We also find new conditions for Riemannian maps to be totally geodesic and harmonic. In section 5, we study slant Riemannian maps as a generalization of slant submersion, give examples, and obtain the harmonicity of such maps. We also find new necessary and sufficient conditions for such maps to be totally geodesic. Moreover, we obtain a decomposition theorem by slant Riemannian maps. In section 6, we define semi-slant Riemannian maps and give examples. In section 7, we introduce hemi-slant Riemannian maps as a generalization of hemi-slant submersions and slant Riemannian maps, give examples, obtain integrability conditions for distribution, and investigate the geometry of these distributions. We also obtain conditions for such maps to be harmonic and totally geodesic, respectively.

Differences in neural crest sensitivity to ethanol account for the infrequency of anterior segment defects in the eye compared with craniofacial anomalies in a zebrafish model of fetal alcohol syndrome.

Ethanol (ETOH) exposure during pregnancy is associated with craniofacial and neurologic abnormalities, but infrequently disrupts the anterior segment of the eye. In these studies, we used zebrafish to investigate differences in the teratogenic effect of ETOH on craniofacial, periocular, and ocular neural crest. Zebrafish eye and neural crest development was analyzed by means of live imaging, TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assay, immunostaining, detection of reactive oxygen species, and in situ hybridization. Our studies demonstrated that foxd3-positive neural crest cells in the periocular mesenchyme and developing eye were less sensitive to ETOH than sox10-positive craniofacial neural crest cells that form the pharyngeal arches and jaw. ETOH increased apoptosis in the retina, but did not affect survival of periocular and ocular neural crest cells. ETOH also did not increase reactive oxygen species within the eye. In contrast, ETOH increased ventral neural crest apoptosis and reactive oxygen species production in the facial mesenchyme. In the eye and craniofacial region, sod2 showed high levels of expression in the anterior segment and in the setting of Sod2 knockdown, low levels of ETOH decreased migration of foxd3-positive neural crest cells into the developing eye. However, ETOH had minimal effect on the periocular and ocular expression of transcription factors (pitx2 and foxc1) that regulate anterior segment development. Neural crest cells contributing to the anterior segment of the eye exhibit increased ability to withstand ETOH-induced oxidative stress and apoptosis. These studies explain the rarity of anterior segment dysgenesis despite the frequent craniofacial abnormalities in fetal alcohol syndrome. Birth Defects Research 109:1212-1227, 2017. © 2017 Wiley Periodicals, Inc.

Embryonic exposure to ethanol reduces cholesterol levels in a chick embryo model of Fetal Alcohol Syndrome

Embryonic exposure to ethanol reduces cholesterol levels in a chick embryo model of Fetal Alcohol Syndrome

Modeling Symptoms of Attention-Deficit Hyperactivity Disorder in a Rat Model of Fetal Alcohol Syndrome.

Several studies indicate the similarity between the symptoms of fetal alcohol syndrome and attention-deficit hyperactivity disorder (ADHD). This study hypothesized that prenatal exposure to ethanol (EtOH) can be used as an animal model of ADHD in Wistar rats. At the first stage of the study, alcohol was delivered to the pregnant dams (237-252 g) by intra-gastric route throughout Gestation Days 8-20 at a dose of 6 g/kg/day. Untreated control group with isocaloric sucrose intubation was also included. Of the 16 male pups (174-180 g), 8 were in the fetal alcohol effects (FAE) group and 8 were in the untreated control group. Subjects went through behavior shaping, discrimination learning and reversal learning. Number of sessions to learn the tasks, response frequency to inhibitory (S-) and excitatory (S+) stimulus features, response latency and inter-response time (IRT) were measured. Significant differences were obtained on only the reversal task. Rats with FAE needed greater number of sessions to learn the reversal task, and they had a higher frequency of incorrect responses in specifically the latter part of the sessions. Our results suggest that reversal learning of FAE rats exhibits deficit in the inhibition of pre-learned responses. Responses behaviorally mimicked attention deficit and impulsivity symptoms of human ADHD. However, the experimental design of the study was not conducive to hyperactivity. Accordingly, rats with FEA can be an alternative to other models since it is not, for example, based on a symptom that is atypical (such as hypertension) to ADHD. Significant difference was obtained in a reversal task between male rats prenatally exposed to ethanol and matched controls. The greater number of sessions for learning and higher frequency of incorrect responses behaviorally mimicked symptoms of ADHD, suggesting that rats with fetal ethanol effects can serve as a useful animal model.

Using optical coherence tomography to rapidly phenotype and quantify congenital heart defects associated with prenatal alcohol exposure.

The most commonly used method to analyze congenital heart defects involves serial sectioning and histology. However, this is often a time-consuming process where the quantification of cardiac defects can be difficult due to problems with accurate section registration. Here we demonstrate the advantages of using optical coherence tomography, a comparatively new and rising technology, to phenotype avian embryo hearts in a model of fetal alcohol syndrome where a binge-like quantity of alcohol/ethanol was introduced at gastrulation. The rapid, consistent imaging protocols allowed for the immediate identification of cardiac anomalies, including ventricular septal defects and misaligned/missing vessels. Interventricular septum thicknesses and vessel diameters for three of the five outflow arteries were also significantly reduced. Outflow and atrioventricular valves were segmented using image processing software and had significantly reduced volumes compared to controls. This is the first study to our knowledge that has 3D reconstructed the late-stage cardiac valves in precise detail to examine their morphology and dimensions. We believe, therefore, that optical coherence tomography, with its ability to rapidly image and quantify tiny embryonic structures in high resolution, will serve as an excellent and cost-effective preliminary screening tool for developmental biologists working with a variety of experimental/disease models.

Heat shock factor 2 is a stress-responsive mediator of neuronal migration defects in models of fetal alcohol syndrome.

Fetal alcohol spectrum disorder (FASD) is a frequent cause of mental retardation. However, the molecular mechanisms underlying brain development defects induced by maternal alcohol consumption during pregnancy are unclear. We used normal and Hsf2-deficient mice and cell systems to uncover a pivotal role for heat shock factor 2 (HSF2) in radial neuronal migration defects in the cortex, a hallmark of fetal alcohol exposure. Upon fetal alcohol exposure, HSF2 is essential for the triggering of HSF1 activation, which is accompanied by distinctive post-translational modifications, and HSF2 steers the formation of atypical alcohol-specific HSF1–HSF2 heterocomplexes. This perturbs the in vivo binding of HSF2 to heat shock elements (HSEs) in genes that control neuronal migration in normal conditions, such as p35 or the MAPs (microtubule-associated proteins, such as Dclk1 and Dcx), and alters their expression. In the absence of HSF2, migration defects as well as alterations in gene expression are reduced. Thus, HSF2, as a sensor for alcohol stress in the fetal brain, acts as a mediator of the neuronal migration defects associated with FASD.Subject Categories Development & Differentiation; Neuroscience

The role of NADPH oxidase in a mouse model of fetal alcohol syndrome

Fetal alcohol syndrome (FAS) is the most common cause of nongenetic mental retardation. Oxidative stress is one of the purported mechanisms. Nicotinamide adenine dinucleotide phosphate oxidase (NOX) is an enzyme involved in the production of reactive oxygen species. Our objective was to evaluate NOX in the fetal brain of a well-validated mouse model of FAS. Timed, pregnant C57BL/6J mice were injected intraperitoneally with 0.03 mL/g of either 25% ethyl alcohol or saline. Fetal brain, liver, and placenta were harvested on gestational day 18. The unit of analysis was the litter; tissue from 6-8 litters in the alcohol and control group was isolated. Evaluation of messenger ribonucleic acid (mRNA) expression of NOX subunits (DUOX1, DUOX2, NOX1, NOX2, NOX3, NOX4, NOXA1, NOXO1, RAC1, p22phox, and p67phox) was performed using quantitative real-time polymerase chain reaction; alcohol vs placebo groups were compared using a Student t test or a Mann-Whitney test (P < .05). Alcohol exposed fetal brains showed significant up-regulation in subunits DUOX2 (1.61 ± 0.28 vs 0.84 ± 0.09; P = .03), NOXA1 (1.75 ± 0.27 vs 1.09 ± 0.06; P = .04), and NOXO1 (1.59 ± 0.10 vs 1.28 ± 0.05; P = .02). Differences in mRNA expression in the placenta were not significant; p67phox was significantly up-regulated in alcohol-exposed livers. Various NOX subunits are up-regulated in fetal brains exposed to alcohol. This effect was not observed in the fetal liver or placenta. Given the available evidence, the NOX system may be involved in the causation of FAS through the generation of reactive oxygen species and may be a potential target for preventative treatment in FAS.

Caspase‐generated neoepitopes as markers of axonal degeneration in neural development and injury

Neuronal death, axonal degeneration, and synapse loss are hallmarks of central nervous system development and injury. Commonly employed markers of apoptosis are TUNEL and immunohistochemistry for activated caspases, but these often fail to report axonal degeneration associated with somatic apoptosis. Axonal degeneration is classically detected by silver degeneration stains, but these have limitations (non‐selectivity, minimal reproducibility and incompatibility with co‐labeling). We demonstrate that antibodies to caspase‐cleaved protein substrates beta‐actin (cleaved by caspase 3 at D244) and alpha‐tubulin (cleaved by caspase 6 at D438) enable superior visualization of neurite degeneration. These antibodies label apoptotic cell bodies and neurites during embryonic and postnatal developmental pruning. In addition, they reveal injured neurons and neurites after induction of apoptosis in a mouse model of fetal alcohol syndrome and in sympathetic neuron cultures induced to undergo apoptosis after nerve growth factor withdrawal. Finally, the markers may have clinical utility, as they highlight degenerating neurites in neonatal hypoxic/ischemic injury and in adult patients with multiple sclerosis. Ongoing work aims to identify novel compartment‐specific neoepitopes that will allow monitoring of somatodendritic, axonal, and synaptic remodeling and degeneration. Supported by NS075869.

Neuroprotective peptides influence cytokine and chemokine alterations in a model of fetal alcohol syndrome

Fetal alcohol syndrome (FAS) is associated with intellectual disability and neurodevelopmental abnormalities. Neuroprotective peptides NAPVSIPQ (NAP) and SALLRSIPA (SAL) can prevent some of the alcohol-induced teratogenesis including fetal death, growth abnormalities, and learning impairment in part by preventing alcohol-induced alterations in N-methyl-D-aspartate receptor gene expression in a mouse model for FAS. We evaluated a panel of cytokines and chemokines to determine whether NAP plus SAL work through a cytokine/chemokine-mediated pathway in preventing these alterations. Using a well-characterized FAS model, timed, pregnant C57BL6/J mice were treated on gestational day (E) 8 with alcohol (0.03 mL/g), placebo, or alcohol plus peptides. Embryos were evaluated at 2 time points: after 6 hours and 10 days later at E18. A panel of cytokines/chemokines was measured using a microsphere-based multiplex immunoassay (Luminex xMAP; Millipore, Billerica, MA). Statistical analysis included Kruskal-Wallis, with P < .05 considered significant. Six hours after treatment, interleukin (IL)-6 and keratinocyte chemoattractant cytokine (KC) were not detectable in the control embryos. Alcohol treatment resulted in detectable levels and significant increases in IL-6 (median, 15.7; range, 10.1-45.9 pg/mL) and KC (median, 45.9; range, 32.5-99.1 pg/mL). Embryos exposed to alcohol plus NAP plus SAL had undetectable IL-6 and KC (both P < .003), similar to the controls. Alcohol exposure resulted in a significant increase of granulocyte colony-stimulating factor (G-CSF) (P < .003) as compared with controls, and treatment with NAP plus SAL prevented the alcohol-induced increase. IL-13 and IL-1β were decreased 6 hours after alcohol exposure, and exposure to alcohol plus NAP plus SAL did not completely ameliorate the decrease. At E18, 10 days after exposure, these alterations were no longer present. Several analytes (regulated upon activation, normal T cell expressed, and secreted, tumor necrosis factor-α, interferon-γ, and IL-4) were not detectable at either time point in any of the groups. Prenatal alcohol exposure acutely results in a significant elevation of IL-6, G-CSF and the KC, which are known to affect N-methyl-D-aspartate receptors. NAP plus SAL treatment prevented alcohol-induced increases. This provides additional insight into the mechanism of alcohol damage in FAS and NAP plus SAL prevention of neurodevelopmental anomalies.

The expression of antioxidant enzymes in a mouse model of fetal alcohol syndrome

Superoxide dismutase, glutathione peroxidase, and catalase prevent cellular damage produced by free radicals. Our objective was to evaluate if prenatal alcohol exposure decreased the expression of antioxidant enzymes in the brain, liver, or placenta of fetal mice. Timed, pregnant C57BL6/J mice were treated on gestational day 8 with intraperitoneal injection of alcohol (0.03 mL/g) or saline (control). Fetuses were harvested on gestational day 18. Fetal brain, liver, and placenta were analyzed for mRNA expression of superoxide dismutase, glutathione peroxidase, and catalase by real-time polymerase chain reaction, with 18S RNA used as reference. Superoxide dismutase, glutathione peroxidase, and catalase expression was lower in fetal brains exposed to alcohol with no differences detected in the liver or placenta between the 2 groups. Maternal alcohol consumption causes a decrease in superoxide dismutase, glutathione peroxidase, and catalase expression in the fetal brain. This may explain the long-term neurologic findings in fetal alcohol syndrome.

43: The role of cell signaling mechanisms in the downregulation of antioxidant enzymes in the fetal brain in a mouse model of fetal alcohol syndrome

downregulation of antioxidant enzymes in the fetal brain in a mouse model of fetal alcohol syndrome Nathan Drever, Talar Kechichian, Nkechi Okonkwo, Debdeep Banerjee, Monica Longo, George Saade, Egle Bytautiene The University of Texas Medical Branch, Obstetrics & Gynecology, Galveston, TX OBJECTIVE: Fetal alcohol syndrome (FAS) is the most common non genetic cause of mental retardation. Prenatal alcohol exposure decreases antioxidant enzymes in the fetal brain in a mouse model of fetal alcohol syndrome. Exposure to alcohol is known to decrease expression of Glycogen Synthase Kinase-3 (GSK-3 ) a serine threonine kinase that regulates glycogen synthesis through phosphorylation and inactivation of glycogen synthase. In addition to regulation of glycogen, GSK-3 is positively correlated with expression of nuclear transcription factors, NFand Nrf2, which are inactive in the cytosol and active within nucleus. Our objective was to evaluate if the effect of prenatal alcohol exposure on fetal brain antioxidants can be explained by an effect on GSK-3 , NFand Nrf2. STUDY DESIGN: A well-characterized FAS model was used (Webster, 1980). Timed, pregnant C57BL6/J mice were treated on gestational day 8 (E8) with alcohol (0.03 mL/g) or vehicle. Fetuses were harvested on gestational day 18 (E18), and the brain prepared for protein analysis. Western Blots were performed assessing the protein expression of GSK-3 and cytosolic and nuclear expression of NFand Nrf2 normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Statistical analysis was performed using Student t-test (significance: p 0.05). RESULTS: GSK-3 protein levels were significantly lower (0.73 vs 1.06 ratio protein/GAPDH; p 0.01) and cytosolic Nrf2 protein levels were significantly higher (0.72 vs 0.43 ratio protein/GAPDH; p 0.01) in the brain of pups exposed to alcohol compared with control. There was no significant difference in cytosolic expression of NFor nuclear expression of NFand Nrf2 between the two groups. CONCLUSION: In utero exposure to alcohol reduces GSK-3 and increases cytosolic Nrf2 protein levels in the fetal brain. These findings provide a mechanism for the decrease in antioxidants in the brain of FAS offspring, and a potential target for preventive strategies. Oral Plenary Session II General www.AJOG.org

431: Protein expression and enzymatic activity of superoxide dismutase in the fetal brain in a mouse model of fetal alcohol syndrome

fetuses with enlarged bladder, bilateral hydronephrosis and normal amniotic fluid volume, diagnosed and enrolled between 20-24 weeks gestation. Exclusion criteria included cystic renal parenchyma, other anomalies, multiple gestation, membrane separation, no karyotype, and cervical length of 1.5cm. Ultrasound evaluation of AFV, renal and bladder dimensions were obtained at 24, 28 and 32 weeks. Delivery and neonatal discharge data were collected. Prospective outcome data at 12 and 24 months is currently being collected. RESULTS: Twenty-seven patients have enrolled. Mean GA at enrollment was 22.7 weeks. Mean AFV at enrollment was 15.9cm (9.5-20.1). One patient underwent TOP. One patient delivered at 27 weeks with a NND at DOL 24 secondary to complications of prematurity. Eight patients remain undelivered. Mean AFV at 32 weeks was 11.6cm (026.7). Delivery data was available for 18 patients. Mean GA at delivery was 39.1 weeks (26.6-40.4). Mean BW was 2910g (1000-3735). CD was performed primarily for fetal malpresentation or history of prior CD. 43% (6/14) demonstrated a normal neonatal serum Cr, while 57% (8/14) demonstrated abnormal neonatal Cr at the time of discharge. Postnatal diagnosis was available for 16 neonates: PUV 50% (9/18) and TRIAD 38.8% (7/18). 55.5% (10/18) neonates required initial respiratory support, but all were discharged on room air. Spontaneous voiding was documented in 55.5% (10/18) neonates at discharge. CONCLUSION: Fetuses with prenatally diagnosed obstructive uropathy and normal AFV are not at increased risk for preterm delivery as reported in LUTO with oligohydramnios. Respiratory and renal functions appear to be preserved in the majority of affected neonates at time of discharge.

Novel approaches in a rat model for fetal alcohol syndrome (fas): Micropet and craniofacial anomalies

Novel approaches in a rat model for fetal alcohol syndrome (fas): Micropet and craniofacial anomalies

Calcium-mediated repression of β-catenin and its transcriptional signaling mediates neural crest cell death in an avian model of fetal alcohol syndrome

Fetal alcohol syndrome (FAS) is a common birth defect in many societies. Affected individuals have neurodevelopmental disabilities and a distinctive craniofacial dysmorphology. These latter deficits originate during early development from the ethanol-mediated apoptotic depletion of cranial facial progenitors, a population known as the neural crest. We showed previously that this apoptosis is caused because acute ethanol exposure activates G-protein-dependent intracellular calcium within cranial neural crest progenitors, and this calcium transient initiates the cell death. The dysregulated signals that reside downstream of ethanol's calcium transient and effect neural crest death are unknown. Here we show that ethanol's repression of the transcriptional effector β-catenin causes the neural crest losses. Clinically relevant ethanol concentrations (22-78 mM) rapidly deplete nuclear β-catenin from neural crest progenitors, with accompanying losses of β-catenin transcriptional activity and downstream genes that govern neural crest induction, expansion, and survival. Using forced expression studies, we show that β-catenin loss of function (via dominant-negative T cell transcription factor [TCF]) recapitulates ethanol's effects on neural crest apoptosis, whereas β-catenin gain-of-function in ethanol's presence preserves neural crest survival. Blockade of ethanol's calcium transient using Bapta-AM normalizes β-catenin activity and prevents the neural crest losses, whereas ionomycin treatment is sufficient to destabilize β-catenin. We propose that ethanol's repression of β-catenin causes the neural crest losses in this model of FAS. β-Catenin is a novel target for ethanol's teratogenicity. β-Catenin/Wnt signals participate in many developmental events and its rapid and persistent dysregulation by ethanol may explain why the latter is such a potent teratogen.

May 2011 DMM Podcast

This podcast features comments by Gerard Evan and others from a recent workshop entitled ‘Cancer as a Microevolutionary Process’, an interview with Frances Balkwill and details on a new Drosophila model of fetal alcohol syndrome from Ulrike Heberlein’s lab. Narrated by Sarah Allan.