RESUMO
BACKGROUND: The essential nutrient choline provides one-carbon units for metabolite synthesis and epigenetic regulation in tissues including brain. Dietary choline intake is often inadequate, and higher intakes are associated with improved cognitive function. OBJECTIVE: Choline supplements confer cognitive improvement for those diagnosed with fetal alcohol spectrum disorder (FASD), a common set of neurodevelopmental impairments; however, the effect sizes have been modest. In this retrospective analysis, we report that genetic polymorphisms affecting choline utilization are associated with cognitive improvement following choline intervention. METHODS: Fifty-two children from the upper midwestern United States and diagnosed with FASD, ages 2-5 y, were randomly assigned to receive choline (500 mg/d; n = 26) or placebo (n = 26) for 9 mo, and were genotyped for 384 choline-related single nucleotide polymorphisms (SNPs). Memory and cognition were assessed at enrollment, study terminus, and at 4-y follow-up for a subset. RESULTS: When stratified by intervention (choline vs. placebo), 14-16 SNPs within the cellular choline transporter gene solute carrier family 44 member 1 (SLC44A1) were significantly associated with performance in an elicited imitation sequential memory task, wherein the effect alleles were associated with the greatest pre-/postintervention improvement. Of these, rs3199966 is a structural variant (S644A) and rs2771040 is a single-nucleotide variant within the 3' untranslated region of the plasma membrane isoform. An additive genetic model best explained the genotype associations. Lesser associations were observed for cognitive outcome and polymorphisms in flavin monooxygenase-3 (FMO3), methylenetetrahydrofolate dehydrogenase-1 (MTHFD1), fatty acid desaturase-2 (FADS2), and adiponectin receptor 1 (ADIPOR1). CONCLUSIONS: These SLC44A1 variants were previously associated with greater vulnerability to choline deficiency. Our data potentially support the use of choline supplements to improve cognitive function in individuals diagnosed with FASD who carry these effect alleles. Although these findings require replication in both retrospective and prospective confirmatory trials, they emphasize the need to incorporate similar genetic analyses of choline-related polymorphisms in other FASD-choline trials, and to test for similar associations within the general FASD population. This trial was registered at www.clinicaltrials.gov as NCT01149538.
Assuntos
Antígenos CD/metabolismo , Colina/farmacologia , Suplementos Nutricionais , Transtornos do Espectro Alcoólico Fetal/tratamento farmacológico , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas de Transporte de Cátions Orgânicos/metabolismo , Polimorfismo de Nucleotídeo Único , Administração Oral , Antígenos CD/genética , Pré-Escolar , Colina/administração & dosagem , Cognição , Feminino , Transtornos do Espectro Alcoólico Fetal/genética , Transtornos do Espectro Alcoólico Fetal/patologia , Genótipo , Humanos , Masculino , Proteínas de Transporte de Cátions Orgânicos/genética , Estudos RetrospectivosRESUMO
Alcohol affects multiple neurotransmitter systems, notably the GABAergic system and has been recognised for a long time as particularly damaging during critical stages of brain development. Nevertheless, data from the literature are most often derived from animal or in vitro models. In order to study the production, migration and cortical density disturbances of GABAergic interneurons upon prenatal alcohol exposure, we performed immunohistochemical studies by means of the proliferation marker Ki67, GABA and calretinin antibodies in the frontal cortical plate of 17 foetal and infant brains antenatally exposed to alcohol, aged 15 weeks' gestation to 22 postnatal months and in the ganglionic eminences and the subventricular zone of the dorsal telencephalon until their regression, i.e., 34 weeks' gestation. Results were compared with those obtained in 17 control brains aged 14 weeks of gestation to 35 postnatal months. We also focused on interneuron vascular migration along the cortical microvessels by confocal microscopy with double immunolabellings using Glut1, GABA and calretinin. Semi-quantitative and quantitative analyses of GABAergic and calretininergic interneuron density allowed us to identify an insufficient and delayed production of GABAergic interneurons in the ganglionic eminences during the two first trimesters of the pregnancy and a delayed incorporation into the laminar structures of the frontal cortex. Moreover, a mispositioning of GABAergic and calretininergic interneurons persisted throughout the foetal life, these cells being located in the deep layers instead of the superficial layers II and III. Moreover, vascular migration of calretininergic interneurons within the cortical plate was impaired, as reflected by low numbers of interneurons observed close to the cortical perforating vessel walls that may in part explain their abnormal intracortical distribution. Our results are globally concordant with those previously obtained in mouse models, in which alcohol has been shown to induce an interneuronopathy by affecting interneuron density and positioning within the cortical plate, and which could account for the neurological disabilities observed in children with foetal alcohol disorder spectrum.
Assuntos
Consumo de Bebidas Alcoólicas , Encéfalo/embriologia , Calbindina 2/metabolismo , Transtornos do Espectro Alcoólico Fetal/metabolismo , Feto/embriologia , Interneurônios/metabolismo , Antígeno Ki-67/metabolismo , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Ácido gama-Aminobutírico/metabolismo , Alcoolismo , Consumo Excessivo de Bebidas Alcoólicas , Encéfalo/metabolismo , Encéfalo/patologia , Estudos de Casos e Controles , Movimento Celular , Feminino , Transtornos do Espectro Alcoólico Fetal/patologia , Feto/metabolismo , Feto/patologia , Lobo Frontal/embriologia , Lobo Frontal/metabolismo , Lobo Frontal/patologia , Neurônios GABAérgicos/metabolismo , Neurônios GABAérgicos/patologia , Humanos , Lactente , Recém-Nascido , Interneurônios/patologia , Masculino , Gravidez , Complicações na Gravidez , Segundo Trimestre da Gravidez , Efeitos Tardios da Exposição Pré-Natal/patologia , Telencéfalo/embriologia , Telencéfalo/metabolismo , Telencéfalo/patologiaRESUMO
Microglia, a type of CNS immune cell, have been shown to contribute to ethanol-activated neuronal death of the stress regulatory proopiomelanocortin (POMC) neuron-producing ß-endorphin peptides in the hypothalamus in a postnatal rat model of fetal alcohol spectrum disorders. We determined whether the microglial extracellular vesicle exosome is involved in the ethanol-induced neuronal death of the ß-endorphin neuron. Extracellular vesicles were prepared from hypothalamic tissues collected from postnatal rats (both males and females) fed daily with 2.5 mg/kg ethanol or control milk formula for 5 d or from hypothalamic microglia cells obtained from postnatal rats, grown in cultures for several days, and then challenged with ethanol or vehicle for 24 h. Nanoparticle tracking analysis and transmission electron microscopy indicated that these vesicles had the size range and shape of exosomes. Ethanol treatments increased the number and the ß-endorphin neuronal killing activity of microglial exosomes both in vivo and in vitro Proteomics analyses of exosomes of cultured microglial cells identified a large number of proteins, including various complements, which were elevated following ethanol treatment. Proteomics data involving complements were reconfirmed using quantitative protein assays. Ethanol treatments also increased deposition of the complement protein C1q in ß-endorphin neuronal cells in both in vitro and in vivo systems. Recombinant C1q protein increased while C1q blockers reduced ethanol-induced C3a/b, C4, and membrane attack complex/C5b9 formations; ROS production; and ultimately cellular death of ß-endorphin neurons. These data suggest that the complement system involving C1q-C3-C4-membrane attack complex and ROS regulates exosome-mediated, ethanol-induced ß-endorphin neuronal death.SIGNIFICANCE STATEMENT Neurotoxic action of alcohol during the developmental period is recognized for its involvement in fetal alcohol spectrum disorders, but the lack of clear understanding of the mechanism of alcohol action has delayed the progress in therapeutic intervention of this disease. Proopiomelanocortin neurons known to regulate stress, energy homeostasis, and immune functions are reported to be killed by developmental alcohol exposure because of activation of microglial immune cells in the brain. While microglia are known to use extracellular vesicles to communicate with neurons for maintaining homeostasis, we show here that ethanol exposure during the developmental period hijacks this system to spread apoptotic factors, including complement protein C1q, to induce the membrane attack complex and reactive super-oxygen species for proopiomelanocortin neuronal killing.
Assuntos
Depressores do Sistema Nervoso Central/farmacologia , Complemento C1q/farmacologia , Etanol/farmacologia , Exossomos/efeitos dos fármacos , Transtornos do Espectro Alcoólico Fetal/patologia , Microglia/efeitos dos fármacos , Pró-Opiomelanocortina/genética , Animais , Animais Recém-Nascidos , Morte Celular/efeitos dos fármacos , Células Cultivadas , Feminino , Transtornos do Espectro Alcoólico Fetal/metabolismo , Hipotálamo/metabolismo , Hipotálamo/patologia , Masculino , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Gravidez , Proteômica , Ratos , Ratos Sprague-Dawley , beta-Endorfina/metabolismoRESUMO
Alcohol exposure during development produces physical and mental abnormalities in the foetus that result in long-term molecular adjustments in the brain, which could underlie the neurobehavioural deficits observed in individuals suffering from foetal alcohol spectrum disorders. In this study, we assessed the effects of curcumin on cognitive impairments caused by prenatal and lactational alcohol exposure (PLAE). Furthermore, we examined whether curcumin could counteract the molecular alterations that may underlie these behavioural impairments. We focused on inflammatory and epigenetic mechanisms by analysing the expression of pro-inflammatory mediators, such as IL-6, TNF-α, and NF-κB, in the hippocampus and prefrontal cortex, as well as microglia and astrocyte activation in the dentate gyrus. We also assessed the activity of histone acetyltransferase in these brain areas. To model binge alcohol drinking, we exposed pregnant C57BL/6 mice to a 20% v/v alcohol solution during gestation and lactation, with limited access periods. We treated male offspring with curcumin during postnatal days (PD28-35) and then evaluated their behaviour in adulthood (PD60). Our results showed that curcumin treatment during the peri-adolescence period improved the anxiety and memory deficits observed in PLAE mice. At the molecular level, we found enhanced histone acetyltransferase activity in mice subjected to PLAE that curcumin treatment could not reverse to baseline levels. These mice also showed increased expression of pro-inflammatory mediators, which could be rescued by curcumin treatment. They also displayed astrogliosis and microglia activation. Our study provides further evidence to support the use of curcumin as a therapeutic agent for counteracting behavioural and molecular alterations induced by PLAE.
Assuntos
Curcumina/uso terapêutico , Modelos Animais de Doenças , Etanol/toxicidade , Transtornos do Espectro Alcoólico Fetal/tratamento farmacológico , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Anti-Inflamatórios não Esteroides/uso terapêutico , Curcumina/farmacologia , Etanol/administração & dosagem , Feminino , Transtornos do Espectro Alcoólico Fetal/patologia , Transtornos do Espectro Alcoólico Fetal/psicologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Aprendizagem em Labirinto/fisiologia , Camundongos , Camundongos Endogâmicos C57BL , Gravidez , Resultado do TratamentoRESUMO
Maternal consumption of alcohol during pregnancy can generate a multitude of deficits in the offspring. Fetal Alcohol Spectrum Disorders, or FASD, describe a palette of potentially life-long phenotypes that result from exposure to ethanol during human gestation. There is no cure for FASD and cognitive-behavioral therapies typically have low success rates, especially in severe cases. The neocortex, responsible for complex cognitive and behavioral function, is altered by prenatal ethanol exposure (PrEE). Supplementation with choline, an essential nutrient, during the prenatal ethanol insult has been associated with a reduction of negative outcomes associated with PrEE. However, choline's ability to prevent deficits within the developing neocortex, as well as the underlying mechanisms, remain unclear. Here, we exposed pregnant mice to 25% ethanol in addition to a 642 mg/L choline chloride supplement throughout gestation to determine the impact of choline supplementation on neocortical and behavioral development in ethanol-exposed offspring. We found that concurrent choline supplementation prevented gross developmental abnormalities associated with PrEE including reduced body weight, brain weight, and cortical length as well as partially ameliorated PrEE-induced abnormalities in intraneocortical circuitry. Additionally, choline supplementation prevented altered expression of RZRß and Id2, two genes implicated in postmitotic patterning of neocortex, and global DNA hypomethylation within developing neocortex. Lastly, choline supplementation prevented sensorimotor behavioral dysfunction and partially ameliorated increased anxiety-like behavior observed in PrEE mice, as assessed by the Suok and Ledge tests. Our results suggest that choline supplementation may represent a potent preventative measure for the adverse outcomes associated with PrEE.
Assuntos
Colina/administração & dosagem , Suplementos Nutricionais , Etanol/toxicidade , Transtornos do Espectro Alcoólico Fetal/tratamento farmacológico , Neocórtex/efeitos dos fármacos , Fenótipo , Animais , Animais Recém-Nascidos , Ansiedade/induzido quimicamente , Ansiedade/tratamento farmacológico , Ansiedade/patologia , Etanol/administração & dosagem , Feminino , Transtornos do Espectro Alcoólico Fetal/patologia , Masculino , Camundongos , Neocórtex/metabolismo , Neocórtex/patologia , GravidezRESUMO
Fetal alcohol spectrum disorder caused by prenatal alcohol exposure includes ocular abnormalities (microphthalmia, photoreceptor dysfunction, cataracts). Zebrafish embryos exposed to ethanol from gastrulation through somitogenesis show severe ocular defects, including microphthalmia and photoreceptor differentiation defects. Ethanol-treated zebrafish had an enlarged ciliary marginal zone (CMZ) relative to the retina size and reduced Müller glial cells (MGCs). Ethanol exposure produced immature photoreceptors with increased proliferation, indicating cell cycle exit failure. Signaling mechanisms in the CMZ were affected by embryonic ethanol exposure, including Wnt signaling in the CMZ, Notch signaling and neurod gene expression. Retinoic acid or folic acid co-supplementation with ethanol rescued Wnt signaling and retinal differentiation. Activating Wnt signaling using GSK3 inhibitor (LSN 2105786; Eli Lilly and Co.) restored retinal cell differentiation pathways. Ethanol exposed embryos were treated with Wnt agonist, which rescued Wnt-active cells in the CMZ, Notch-active cells in the retina, proliferation, and photoreceptor terminal differentiation. Our results illustrate the critical role of Wnt signaling in ethanol-induced retinal defects.
Assuntos
Etanol/efeitos adversos , Transtornos do Espectro Alcoólico Fetal/metabolismo , Retina/patologia , Doenças Retinianas/metabolismo , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Diferenciação Celular , Proliferação de Células/efeitos dos fármacos , Modelos Animais de Doenças , Embrião não Mamífero/efeitos dos fármacos , Feminino , Transtornos do Espectro Alcoólico Fetal/patologia , Ácido Fólico/farmacologia , Proteínas do Tecido Nervoso/genética , Células Fotorreceptoras de Vertebrados/citologia , Células Fotorreceptoras de Vertebrados/efeitos dos fármacos , Células Fotorreceptoras de Vertebrados/metabolismo , Gravidez , Receptores Notch/metabolismo , Retina/efeitos dos fármacos , Retina/metabolismo , Doenças Retinianas/induzido quimicamente , Doenças Retinianas/patologia , Tretinoína/farmacologia , Via de Sinalização Wnt , Peixe-Zebra/embriologia , Peixe-Zebra/genéticaRESUMO
There are disorders in children, covered by the umbrella term "fetal alcohol spectrum disorder" (FASD), that occur as result of alcohol consumption during pregnancy and lactation. They appear, at least in part, to be related to the oxidative stress generated by ethanol. Ethanol metabolism generates reactive oxygen species and depletes the antioxidant molecule glutathione (GSH), leading to oxidative stress and lipid and protein damage, which are related to growth retardation and neurotoxicity, thereby increasing the incidence of FASD. Furthermore, prenatal and postnatal exposure to ethanol in dams, as well as increasing oxidation in offspring, causes malnutrition of several micronutrients such as the antioxidant folic acid and selenium (Se), affecting their metabolism and bodily distribution. Although abstinence from alcohol is the only way to prevent FASD, it is possible to reduce its harmful effects with a maternal dietary antioxidant therapy. In this review, folic acid and Se have been chosen to be analyzed as antioxidant intervention systems related to FASD because, like ethanol, they act on the methionine metabolic cycle, being related to the endogenous antioxidants GSH and glutathione peroxidase. Moreover, several birth defects are related to poor folate and Se status.
Assuntos
Etanol/efeitos adversos , Transtornos do Espectro Alcoólico Fetal , Ácido Fólico/uso terapêutico , Estresse Oxidativo/efeitos dos fármacos , Efeitos Tardios da Exposição Pré-Natal , Selênio/uso terapêutico , Animais , Criança , Pré-Escolar , Feminino , Transtornos do Espectro Alcoólico Fetal/metabolismo , Transtornos do Espectro Alcoólico Fetal/patologia , Transtornos do Espectro Alcoólico Fetal/prevenção & controle , Humanos , Masculino , Micronutrientes/uso terapêutico , Gravidez , Efeitos Tardios da Exposição Pré-Natal/metabolismo , Efeitos Tardios da Exposição Pré-Natal/patologia , Efeitos Tardios da Exposição Pré-Natal/prevenção & controleRESUMO
Ethanol is the most important teratogen agent in humans. Prenatal alcohol exposure can lead to a wide range of adverse effects, which are broadly termed as fetal alcohol spectrum disorder (FASD). The most severe consequence of maternal alcohol abuse is the development of fetal alcohol syndrome, defined by growth retardation, facial malformations, and central nervous system impairment expressed as microcephaly and neurodevelopment abnormalities. These alterations generate a broad range of cognitive abnormalities such as learning disabilities and hyperactivity and behavioural problems. Socioeconomic status, ethnicity, differences in genetic susceptibility related to ethanol metabolism, alcohol consumption patterns, obstetric problems, and environmental influences like maternal nutrition, stress, and other co-administered drugs are all factors that may influence FASD manifestations. Recently, much attention has been paid to the role of nutrition as a protective factor against alcohol teratogenicity. There are a great number of papers related to nutritional treatment of nutritional deficits due to several factors associated with maternal consumption of alcohol and with eating and social disorders in FASD children. Although research showed the clinical benefits of nutritional interventions, most of work was in animal models, in a preclinical phase, or in the prenatal period. However, a minimum number of studies refer to postnatal nutrition treatment of neurodevelopmental deficits. Nutritional supplementation in children with FASD has a dual objective: to overcome nutritional deficiencies and to reverse or improve the cognitive deleterious effects of prenatal alcohol exposure. Further research is necessary to confirm positive results, to determine optimal amounts of nutrients needed in supplementation, and to investigate the collective effects of simultaneous multiple-nutrient supplementation.
Assuntos
Transtornos do Espectro Alcoólico Fetal/dietoterapia , Transtornos Neurocognitivos/dietoterapia , Consumo de Bebidas Alcoólicas/efeitos adversos , Consumo de Bebidas Alcoólicas/metabolismo , Animais , Etanol/efeitos adversos , Etanol/metabolismo , Transtornos do Espectro Alcoólico Fetal/genética , Transtornos do Espectro Alcoólico Fetal/metabolismo , Transtornos do Espectro Alcoólico Fetal/patologia , Predisposição Genética para Doença , Humanos , Transtornos Neurocognitivos/genética , Transtornos Neurocognitivos/metabolismo , Transtornos Neurocognitivos/patologiaRESUMO
Importance: Children who receive a diagnosis of fetal alcohol spectrum disorder may have a characteristic facial appearance in addition to neurodevelopmental impairment. It is not well understood whether there is a gradient of facial characteristics of children who did not receive a diagnosis of fetal alcohol spectrum disorder but who were exposed to a range of common drinking patterns during pregnancy. Objective: To examine the association between dose, frequency, and timing of prenatal alcohol exposure and craniofacial phenotype in 12-month-old children. Design, Setting, and Participants: A prospective cohort study was performed from January 1, 2011, to December 30, 2014, among mothers recruited in the first trimester of pregnancy from low-risk, public maternity clinics in metropolitan Melbourne, Australia. A total of 415 white children were included in this analysis of 3-dimensional craniofacial images taken at 12 months of age. Analysis was performed with objective, holistic craniofacial phenotyping using dense surface models of the face and head. Partial least square regression models included covariates known to affect craniofacial shape. Exposures: Low, moderate to high, or binge-level alcohol exposure in the first trimester or throughout pregnancy. Main Outcomes and Measures: Anatomical differences in global and regional craniofacial shape between children of women who abstained from alcohol during pregnancy and children with varying levels of prenatal alcohol exposure. Results: Of the 415 children in the study (195 girls and 220 boys; mean [SD] age, 363.0 [8.3] days), a consistent association between craniofacial shape and prenatal alcohol exposure was observed at almost any level regardless of whether exposure occurred only in the first trimester or throughout pregnancy. Regions of difference were concentrated around the midface, nose, lips, and eyes. Directional visualization showed that these differences corresponded to general recession of the midface and superior displacement of the nose, especially the tip of the nose, indicating shortening of the nose and upturning of the nose tip. Differences were most pronounced between groups with no exposure and groups with low exposure in the first trimester (forehead), moderate to high exposure in the first trimester (eyes, midface, chin, and parietal region), and binge-level exposure in the first trimester (chin). Conclusions and Relevance: Prenatal alcohol exposure, even at low levels, can influence craniofacial development. Although the clinical significance of these findings is yet to be determined, they support the conclusion that for women who are or may become pregnant, avoiding alcohol is the safest option.
Assuntos
Consumo Excessivo de Bebidas Alcoólicas/patologia , Anormalidades Craniofaciais/induzido quimicamente , Anormalidades Craniofaciais/diagnóstico por imagem , Transtornos do Espectro Alcoólico Fetal/patologia , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Austrália , Estudos de Coortes , Anormalidades Craniofaciais/patologia , Fácies , Feminino , Humanos , Lactente , Masculino , Gravidez , Efeitos Tardios da Exposição Pré-Natal/patologia , Prevalência , Estudos Prospectivos , Crânio/anormalidades , Tomografia Computadorizada por Raios XRESUMO
Prenatal alcohol exposure is known to cause fetal growth restriction and disturbances in amino acid bioavailability. Alterations in these parameters can persist into adulthood and low birth weight can lead to altered fetal programming. Glutamine has been associated with the synthesis of other amino acids, an increase in protein synthesis and it is used clinically as a nutrient supplement for low birth weight infants. The aim of this study was to explore the effect of repeated maternal alcohol exposure and L-glutamine supplementation on fetal growth and amino acid bioavailability during the third trimester-equivalent period in an ovine model. Pregnant sheep were randomly assigned to four groups, saline control, alcohol (1.75-2.5 g/kg), glutamine (100 mg/kg, three times daily) or alcohol + glutamine. In this study, a weekend binge drinking model was followed where treatment was done 3 days per week in succession from gestational day (GD) 109-132 (normal term ~147). Maternal alcohol exposure significantly reduced fetal body weight, height, length, thoracic girth and brain weight, and resulted in decreased amino acid bioavailability in fetal plasma and placental fluids. Maternal glutamine supplementation successfully mitigated alcohol-induced fetal growth restriction and improved the bioavailability of glutamine and glutamine-related amino acids such as glycine, arginine, and asparagine in the fetal compartment. All together, these findings show that L-glutamine supplementation enhances amino acid availability in the fetus and prevents alcohol-induced fetal growth restriction.
Assuntos
Suplementos Nutricionais , Transtornos do Espectro Alcoólico Fetal/prevenção & controle , Retardo do Crescimento Fetal/prevenção & controle , Glutamina/farmacologia , Animais , Feminino , Transtornos do Espectro Alcoólico Fetal/metabolismo , Transtornos do Espectro Alcoólico Fetal/patologia , Retardo do Crescimento Fetal/induzido quimicamente , Retardo do Crescimento Fetal/metabolismo , Retardo do Crescimento Fetal/patologia , Gravidez , OvinosRESUMO
Adolescent binge alcohol exposure has long-lasting effects on the expression of hypothalamic genes that regulate the stress response, even in the absence of subsequent adult alcohol exposure. This suggests that alcohol can induce permanent gene expression changes, potentially through epigenetic modifications to specific genes. Epigenetic modifications can be transmitted to future generations therefore, and in these studies we investigated the effects of adolescent binge alcohol exposure on hypothalamic gene expression patterns in the F1 generation offspring. It has been well documented that maternal alcohol exposure during fetal development can have devastating neurological consequences. However, less is known about the consequences of maternal and/or paternal alcohol exposure outside of the gestational time frame. Here, we exposed adolescent male and female rats to a repeated binge EtOH exposure paradigm and then mated them in adulthood. Hypothalamic samples were taken from the offspring of these animals at postnatal day (PND) 7 and subjected to a genome-wide microarray analysis followed by qRT-PCR for selected genes. Importantly, the parents were not intoxicated at the time of mating and were not exposed to EtOH at any time during gestation therefore the offspring were never directly exposed to EtOH. Our results showed that the offspring of alcohol-exposed parents had significant differences compared to offspring from alcohol-naïve parents. Specifically, major differences were observed in the expression of genes that mediate neurogenesis and synaptic plasticity during neurodevelopment, genes important for directing chromatin remodeling, posttranslational modifications or transcription regulation, as well as genes involved in regulation of obesity and reproductive function. These data demonstrate that repeated binge alcohol exposure during pubertal development can potentially have detrimental effects on future offspring even in the absence of direct fetal alcohol exposure.
Assuntos
Intoxicação Alcoólica/genética , Consumo Excessivo de Bebidas Alcoólicas/genética , Etanol/toxicidade , Transtornos do Espectro Alcoólico Fetal/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Hipotálamo/metabolismo , Efeitos Tardios da Exposição Pré-Natal/genética , Intoxicação Alcoólica/embriologia , Intoxicação Alcoólica/patologia , Animais , Consumo Excessivo de Bebidas Alcoólicas/tratamento farmacológico , Consumo Excessivo de Bebidas Alcoólicas/patologia , Biomarcadores/metabolismo , Peso Corporal , Feminino , Transtornos do Espectro Alcoólico Fetal/tratamento farmacológico , Transtornos do Espectro Alcoólico Fetal/patologia , Perfilação da Expressão Gênica , Hipotálamo/efeitos dos fármacos , Hipotálamo/patologia , Masculino , Análise de Sequência com Séries de Oligonucleotídeos , Gravidez , Efeitos Tardios da Exposição Pré-Natal/induzido quimicamente , Efeitos Tardios da Exposição Pré-Natal/patologia , RNA Mensageiro/genética , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
BACKGROUND: The link between the numerous cognitive, motor, and behavioral difficulties of individuals with fetal alcohol spectrum disorders (FASD) and underlying specific structural brain injuries can be investigated using high-resolution imaging. Differential sensitivity of the brain's "relay" stations, namely the deep gray matter structures, may play a key factor given their multifaceted role in brain function. The purpose of our study was to analyze differences in deep gray matter volumes of children and adolescents with FASD relative to age/sex-matched controls and to examine whether any volume differences were consistent across the age range of neurodevelopment. METHODS: Children and adolescents (N = 28, 6 to 17 years) diagnosed with FASD and 56 age- and sex-matched healthy controls (i.e., 2 matched controls per FASD subject) underwent 3-dimensional T1-weighted MRI scans that were used for the automated volume measurement (FreeSurfer) of the intracranial space, total white matter, cortical gray matter, and 6 deep gray matter structures, namely the hippocampus, amygdala, thalamus, caudate, putamen, and globus pallidus, with left and right measured separately. Volumes were compared between FASD and controls, as well as changes with age. RESULTS: Significant reductions of volume in FASD were observed for the intracranial vault (7.6%), total white matter (8.6%), total cortical gray matter (7.8%), and total deep gray matter (13.1%). All 6 deep gray matter structures showed significant volume reductions bilaterally with the caudate (approximately 16%) and globus pallidus (approximately 18%) being most affected. The hippocampus, thalamus, and globus pallidus showed reductions in all 3 age subgroups (6 to 9, 10 to 13, and 14 to 17 years) but the caudate and putamen had smaller volumes for FASD only within the 2 youngest subgroups; the amygdala was only smaller for FASD in the 2 oldest subgroups. CONCLUSIONS: Significant, but variable, volume reductions throughout the deep gray matter are observed over a wide age range of 6 to 17 years in FASD.
Assuntos
Encéfalo/efeitos dos fármacos , Depressores do Sistema Nervoso Central/efeitos adversos , Transtornos Cognitivos/epidemiologia , Etanol/efeitos adversos , Transtornos do Espectro Alcoólico Fetal/epidemiologia , Transtornos do Espectro Alcoólico Fetal/patologia , Adolescente , Tonsila do Cerebelo/patologia , Transtornos de Deficit da Atenção e do Comportamento Disruptivo/epidemiologia , Transtornos de Deficit da Atenção e do Comportamento Disruptivo/patologia , Encéfalo/crescimento & desenvolvimento , Encéfalo/patologia , Núcleo Caudado/patologia , Criança , Transtornos Cognitivos/induzido quimicamente , Transtornos Cognitivos/patologia , Comorbidade , Feminino , Globo Pálido/patologia , Hipocampo/patologia , Humanos , Imageamento por Ressonância Magnética/métodos , Masculino , Fenótipo , Gravidez , Putamen/patologia , Tálamo/patologiaRESUMO
BACKGROUND: Fetal alcohol spectrum disorder (FASD) is a challenging public health problem. Previous studies have found an association between FASD and oxidative stress. In the present study, we assessed the role of oxidative stress in ethanol-induced embryonic damage and the effect of (-)-epigallocatechin-3-gallate (EGCG), a powerful antioxidant extracted from green tea, on the development of FASD in a murine model. METHODS: Pregnant female mice were given intraperitoneal ethanol (25%, 0.005 to 0.02 ml/g) on gestational day 8 (G8) to establish the FASD model. On G10.25, mice were sacrificed and embryos were collected and photographed to determine head length (HL), head width (HW), and crown rump length (CRL). For mice given EGCG, administration was through a feeding tube on G7 and G8 (dose: 200, 300, or 400 mg/kg/d, the total amount for a day was divided into 2 equal portions). G10.25 embryos were evaluated morphologically. Brain tissues of G9.25 embryos were used for RT-PCR and western blotting of neural marker genes and proteins and detection of oxidative stress indicators. RESULTS: Administration of ethanol to pregnant mice on G8 led to the retardation of embryonic growth and down-regulation of neural marker genes. In addition, administration of ethanol (0.02 ml/g) led to the elevation of oxidative stress indicators [hydrogen peroxide (H2O2) and malondialdehyde (MDA)]. Administration of EGCG on G7 and G8 along with ethanol on G8 ameliorated the ethanol-induced growth retardation. Mice given EGCG (400 mg/kg/d) along with ethanol had embryo sizes and neural marker genes expression similar to the normal controls. Furthermore, EGCG (400 mg/kg on G7 and G8) inhibited the increase in H2O2 and MDA. CONCLUSIONS: In a murine model, oxidative stress appears to play an important role in ethanol-induced embryonic growth retardation. EGCG can prevent some of the embryonic injuries caused by ethanol.
Assuntos
Catequina/análogos & derivados , Depressores do Sistema Nervoso Central/antagonistas & inibidores , Depressores do Sistema Nervoso Central/toxicidade , Etanol/antagonistas & inibidores , Etanol/toxicidade , Transtornos do Espectro Alcoólico Fetal/tratamento farmacológico , Fármacos Neuroprotetores/uso terapêutico , Animais , Western Blotting , Encéfalo/embriologia , Encéfalo/patologia , Catequina/uso terapêutico , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Transtornos do Espectro Alcoólico Fetal/patologia , Feto/patologia , Marcadores Genéticos , Humanos , Peróxido de Hidrogênio/toxicidade , Recém-Nascido , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Proteínas do Tecido Nervoso/metabolismo , Estresse Oxidativo/fisiologia , Gravidez , Espécies Reativas de Oxigênio , Reação em Cadeia da Polimerase Via Transcriptase Reversa , CháRESUMO
BACKGROUND: Ethanol is the main addictive and neurotoxic constituent of alcohol. Ethanol exposure during embryonic development causes dysfunction of the central nervous system (CNS) and leads to fetal alcohol spectrum disorders. The cerebellum is one of the CNS regions that are particularly vulnerable to ethanol toxic effects. Retinoic acid (RA) is a physiologically active metabolite of vitamin A that is locally synthesized in the cerebellum. Studies have shown that RA is required for neuronal development, but it remains unknown if ethanol impairs RA signaling and thus induces neuronal malformations. In this study, we tested the hypothesis that ethanol impairs the expression and activation of RA receptors in cerebellum and in cerebellar granule cells. METHODS: The cerebellum of ethanol unexposed and exposed pups was used to study the expression of retinoic acid receptors (RARs or RXRs) by immunohistochemistry and by Western blot analysis. We also studied the effect of ethanol on expression of RA receptors in the cerebellar granule cells. Activation of RA receptors (DNA-binding activities) in response to high-dose ethanol was determined by electrophoretic mobility shift and supershift assays. RESULTS: Findings from these studies demonstrated that ethanol exposure reduced the expression of RARalpha/gamma while it increased the expression of RXRalpha/gamma in the cerebellum and in cerebellar granule neurons. Immuno-histological studies further strengthened the expression pattern of RA receptors in response to ethanol. The DNA-binding activity of RARs was reduced, while DNA-binding activity of RXRs was increased in response to ethanol exposure. CONCLUSION: For the first time, our studies have demonstrated that high-dose ethanol affects the expression and activation of RA receptors, which could impair the signaling events and induce harmful effects on the survival and differentiation of cerebellar granule cells. Taken together, these findings could provide insight into the treatment options for brain defects caused by excessive ethanol exposure, such as in Fetal Alcohol Spectrum Disorders.
Assuntos
Cerebelo/metabolismo , Modelos Animais de Doenças , Etanol/administração & dosagem , Transtornos do Espectro Alcoólico Fetal/metabolismo , Receptores do Ácido Retinoico/antagonistas & inibidores , Receptores do Ácido Retinoico/metabolismo , Consumo de Bebidas Alcoólicas/efeitos adversos , Consumo de Bebidas Alcoólicas/metabolismo , Consumo de Bebidas Alcoólicas/patologia , Animais , Animais Recém-Nascidos , Células Cultivadas , Cerebelo/efeitos dos fármacos , Cerebelo/patologia , Grânulos Citoplasmáticos/efeitos dos fármacos , Grânulos Citoplasmáticos/metabolismo , Feminino , Transtornos do Espectro Alcoólico Fetal/patologia , Gravidez , Distribuição Aleatória , Ratos , Ratos Long-Evans , Receptor alfa de Ácido Retinoico , Receptor gama de Ácido RetinoicoRESUMO
The present study explored the effects of gestational ethanol exposure on enduring changes in the distribution of projection neurons and local circuit neurons in somatosensory/motor cortex. Critical events in corticogenesis occur during macaque gestation: the first six weeks of gestation include the period of primary stem cell production and the next 18 weeks are marked by the birth, migration, early differentiation, and death of cortical neurons. Monkeys were exposed to ethanol (or saline) one day per week during the first six or during the entire 24 weeks of gestation. Offspring were killed as adolescents. Projection neurons and local circuit neurons were identified immunohistochemically with antibodies directed against glutamate and anti-GABA, respectively. In all animals, both projection neurons and local circuit neurons were distributed in all laminae of both somatosensory and motor cortices. Ethanol did not affect the size of Cresyl Violet-stained, glutamate-positive, or GABA-immunolabeled somata, however, it did decrease neuronal density. The total density of Cresyl Violet-stained neurons was reduced in monkeys treated with ethanol (or saline) one day per week during the first six weeks of gestation and during the entire 24 weeks of gestation. Similar reductions were detected for glutamate- and GABA-positive neurons. The densities of Cresyl Violet-stained and of glutamate- and GABA-expressing neurons were reduced in all cortical layers. The only exception was layer V which was unaffected in monkeys treated with ethanol (or saline) one day per week during the first six weeks of gestation and during the entire 24 weeks of gestation. Thus, the parallel effects on both neuronal subpopulations suggest that ethanol targets a population of undetermined neuronal precursors.
Assuntos
Depressores do Sistema Nervoso Central/toxicidade , Etanol/toxicidade , Transtornos do Espectro Alcoólico Fetal/patologia , Ácido Glutâmico/metabolismo , Córtex Motor/metabolismo , Córtex Somatossensorial/metabolismo , Ácido gama-Aminobutírico/metabolismo , Animais , Contagem de Células , Feminino , Imuno-Histoquímica , Macaca nemestrina , Córtex Motor/efeitos dos fármacos , Vias Neurais/citologia , Vias Neurais/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Gravidez , Efeitos Tardios da Exposição Pré-Natal , Córtex Somatossensorial/citologia , Córtex Somatossensorial/efeitos dos fármacosRESUMO
BACKGROUND: Recent research has suggested that oxidative stress is a potential mechanism for alcohol-induced injury and that supplementation with antioxidants can ameliorate alcohol-induced damage. In this study, two known antioxidants, melatonin and U83836E, were assessed for their effectiveness in blocking the expected alcohol-induced cerebellar Purkinje cell loss in neonatal rat pups. METHODS: Sprague-Dawley rat pups were artificially reared from postnatal days (PDs) 4-9 and were exposed to either alcohol or antioxidants (melatonin or U83836E) individually or in combination. A normal control group (raised by rat dams) was included in this study. On PD 9, the brain from each pup was removed and weighed, and the cerebellar vermis was processed for stereological cell counting. RESULTS: Alcohol exposure during the brain growth spurt produced microencephaly, in addition to significant decreases in the number and density of Purkinje cells in lobule I and the volume of lobule I. The antioxidants did not reduce any of the adverse effects observed from alcohol exposure, and they did not decrease the Purkinje cell number when administered alone. Furthermore, antioxidants did not change the only blood alcohol concentration measured on PD 6. CONCLUSIONS: The results confirmed alcohol-induced microencephaly and cerebellar Purkinje cell loss from neonatal alcohol exposure, and they showed that neither antioxidant could attenuate these adverse effects on the developing brain. The inability of antioxidants to reduce Purkinje cell loss from neonatal alcohol exposure suggests the existence of alternative mechanisms for developmental alcohol-induced Purkinje cell loss.
Assuntos
Antioxidantes/farmacologia , Cerebelo/patologia , Cromanos/farmacologia , Etanol/toxicidade , Transtornos do Espectro Alcoólico Fetal/patologia , Melatonina/farmacologia , Piperazinas/farmacologia , Células de Purkinje/patologia , Fatores Etários , Animais , Feminino , Estresse Oxidativo/efeitos dos fármacos , Gravidez , Pré-Medicação , RatosRESUMO
The initial diagnosis of fetal alcohol syndrome (FAS) in the United States was made because of the facial features common to the first cohort of patients. This article reviews the development of an FAS mouse model whose craniofacial features are remarkably similar to those of affected humans. The model is based on short-term maternal treatment with a high dosage of ethanol at stages of pregnancy that are equivalent to Weeks 3 and 4 of human gestation. At these early stages of development, alcohol's insult to the developing face is concurrent with that to the brain, eyes, and inner ear. That facial and central nervous system defects consistent with FAS can be induced by more "realistic" alcohol dosages as illustrated with data from an oral alcohol intake mouse model in which maternal blood alcohol levels do not exceed 200 mg/dl. The ethanol-induced pathogenesis involves apoptosis that occurs within 12 hrs of alcohol exposure in selected cell populations of Day 7, 8, and 9 mouse embryos. Experimental evidence from other species also shows that apoptosis underlies ethanol-induced malformations. With knowledge of sensitive and resistant cell populations at specific developmental stages, studies designed to identify the basis for these differing cellular responses and, therefore, to determine the primary mechanisms of ethanol's teratogenesis are possible. For example, microarray comparisons of sensitive and resistant embryonic cell populations have been made, as have in situ studies of gene expression patterns in the populations of interest. Studies that illustrate agents that are effective in diminishing or exacerbating ethanol's teratogenesis have also been helpful in determining mechanisms. Among these agents are antioxidants, sonic hedgehog protein, retinoids, and the peptides SAL and NAP.
Assuntos
Consumo de Bebidas Alcoólicas/efeitos adversos , Anormalidades Craniofaciais/etiologia , Transtornos do Espectro Alcoólico Fetal/etiologia , Animais , Encéfalo/anormalidades , Encéfalo/efeitos dos fármacos , Encéfalo/ultraestrutura , Anormalidades Craniofaciais/embriologia , Anormalidades Craniofaciais/patologia , Modelos Animais de Doenças , Embrião de Mamíferos/ultraestrutura , Anormalidades do Olho/embriologia , Anormalidades do Olho/etiologia , Feminino , Transtornos do Espectro Alcoólico Fetal/embriologia , Transtornos do Espectro Alcoólico Fetal/patologia , Idade Gestacional , Humanos , Desenvolvimento Maxilofacial/efeitos dos fármacos , Camundongos , GravidezRESUMO
Based on previous in vitro studies that have illustrated prevention of ethanol-induced cell death by antioxidants, using an in vivo model, we have tested the anti-teratogenic potential of a potent synthetic superoxide dismutase plus catalase mimetic, EUK-134. The developing limb of C57BL/6J mice, which is sensitive to ethanol-induced reduction defects, served as the model system. On their ninth day of pregnancy, C57BL/6J mice were administered ethanol (two intraperitoneal doses of 2.9 g/kg given 4 h apart) alone or in combination with EUK-134 (two doses of 10 mg/kg). Pregnant control mice were similarly treated with either vehicle or EUK-134, alone. Within 15 h of the initial ethanol exposure, excessive apoptotic cell death was observed in the apical ectodermal ridge (AER) of the newly forming forelimb buds. Forelimb defects, including postaxial ectrodactyly, metacarpal, and ulnar deficiencies, occurred in 67.3% of the ethanol-exposed fetuses that were examined at 18 days of gestation. The right forelimbs were preferentially affected. No limb malformations were observed in control fetuses. Cell death in the AER of embryos concurrently exposed to ethanol and EUK-134 was notably reduced compared with that in embryos from ethanol-treated dams. Additionally, the antioxidant treatment reduced the incidence of forelimb malformations to 35.9%. This work illustrates that antioxidants can significantly improve the adverse developmental outcome that results from ethanol exposure in utero, diminishing the incidence and severity of major malformations that result from exposure to this important human teratogen.
Assuntos
Antioxidantes/uso terapêutico , Etanol/toxicidade , Transtornos do Espectro Alcoólico Fetal/tratamento farmacológico , Deformidades Congênitas dos Membros/prevenção & controle , Compostos Organometálicos/uso terapêutico , Salicilatos/uso terapêutico , Animais , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Catalase , Avaliação Pré-Clínica de Medicamentos , Feminino , Transtornos do Espectro Alcoólico Fetal/embriologia , Transtornos do Espectro Alcoólico Fetal/patologia , Reabsorção do Feto/induzido quimicamente , Reabsorção do Feto/prevenção & controle , Membro Anterior/anormalidades , Membro Anterior/efeitos dos fármacos , Deformidades Congênitas dos Membros/induzido quimicamente , Deformidades Congênitas dos Membros/embriologia , Troca Materno-Fetal , Metacarpo/anormalidades , Metacarpo/embriologia , Camundongos , Camundongos Endogâmicos C57BL , Compostos Organometálicos/farmacologia , Estresse Oxidativo , Gravidez , Salicilatos/farmacologia , Superóxido Dismutase , Dedos do Pé/anormalidades , Dedos do Pé/embriologia , Ulna/anormalidades , Ulna/embriologiaRESUMO
The thalamo-cortico-thalamic loop was investigated in adult rats exposed to ethanol during the last week of fetal life. Animals underwent either cortical or thalamic injections of lectin-conjugated horseradish peroxidase. Results demonstrate that prenatal exposure to ethanol causes permanent changes in the thalamocortical circuits. Alterations of thalamo-cortical and cortico-thalamic projections are concentrated at the level of axon terminal fields. The most severe thalamic damage is observed in the anterior intralaminar and midline nuclei; crossed cortico-thalamic projections also appear to be severely impaired. In the cortex, the damage to thalamic terminals displays a medio-lateral gradient of increasing severity through sensori-motor areas, with the lateral fields more impaired. Cells of origin of thalamo-cortical and cortico-thalamic projections are less affected by prenatal ethanol exposure: in the thalamus and layer 5 of sensori-motor cortex labeled cells exhibit normal values of areal numeric density. Conversely, cortico-thalamic neurons of layer 6, especially in the lateral agranular sensori-motor field, display smaller values of areal density than those of normal animals. Possible mechanisms underlying the establishment of these abnormalities are discussed.