Ethanol action in Japanese medaka: Alteration in specific gene methylation

日本青鳉中的乙醇作用：特定基因甲基化的改变

• 批准号: 7294529
• 负责人: Asok K Dasmahapatra
• 金额: $ 7万
• 依托单位: UNIVERSITY OF MISSISSIPPI
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-15 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7294529
• 关键词: Adult; Alcohol consumption; Alcohol dehydrogenase; Alcohols; Aldehyde dehydrogenase (NAD+); Animal Model; Animals; Antioxidants; Attenuated; Biological; Biological Factors; Cardiovascular system; Condition; Congenital neurologic anomalies; CpG Islands; DNA; DNA Methylation; Data; Defect; Development; Disease; Embryo; Embryonic Development; Embryonic and Fetal Development; Enzyme Gene; Enzymes; Epigenetic Process; Ethanol; Ethanol Metabolism; Ethanol toxicity; Event; Exhibits; Fetal Alcohol Spectrum Disorder; Fetal Alcohol Syndrome; Fetal Development; Fishes; Foundations; Functional disorder; Future; Gastritis; Gene Expression; Gene Targeting; Genes; Growth; Hepatitis; Human; Incidence; Individual; Japanese Killifish; Kudzu; Limb structure; Malignant Neoplasms; Measures; Messenger RNA; Methylation; Modeling; Modification; Molecular; Morphogenesis; Mothers; NADH; Neuraxis; Newborn Infant; Nicotinamide adenine dinucleotide; Organ; Oxidation-Reduction; Oxidative Stress; Pattern; Phenotype; Pregnancy; Prevention; Preventive; Process; Promoter Regions; Pueraria; Pueraria lobata; Recovery; Research; Retinal; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Skeletal system; Source; Staging; Structure; Supplementation; Techniques; Testing; Therapeutic Agents; Therapeutic Intervention; Tretinoin; Vitamin E; Woman; alcohol effect; alcohol exposure; aldehyde dehydrogenase 1A2; aldehyde dehydrogenases; craniofacial; daidzein; daidzin; egg; experience; gene induction; mRNA Expression; prevent; promoter; puerarin

DESCRIPTION (provided by applicant): The primary focus of the project described herein is to study the effects of ethanol on aldehyde dehydrogenase 1A2 (Aldh1A2) gene expression in Japanese medaka embryogenesis and to determine whether alteration in methylation pattern of the CpG island at the promoter region of this gene is a possible cause of ethanol teratogenesis. Ethanol induces fetal alcohol spectrum disorder (FASD) in the newborn babies of mothers who ingested alcohol during pregnancy. Among FASD, fetal alcohol syndrome (FAS) is the most clinically recognizable form identified by growth retardation, central nervous system (CNS) malformation and dysfunction, and has a distinctive pattern of craniofacial,cardiovascular, and limb defects. The molecular mechanism by which ethanol perturbs fetal development is unknown. Moreover, prevention of FAS, other than women abstaining from drinking alcohol during pregnancy, is not known. Previously, we have demonstrated that medaka embryos exposed to ethanol during embryogenesis have developed precocious FAS features in craniofacial, cardiovascular and skeletal organs which can be compared with FAS features observed in human. These findings prompted us to use medaka as a unique model for searching genes responsible for FAS. Our hypothesis is ethanol metabolism by the enzymes alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) alters cellular NAD + /NADH ratio and induces oxidative stress. This altered redox state is able to induce epigenetic modifications in DNA molecules of specific genes and thus modulate the normal development of the embryo. In this application we will study the DNA methylation pattern in the CpG island in the promoter region of Aldh1A2 gene, which we have identified as an ethanol target gene in medaka embryogenesis. We will also determine the effect of a natural product, kudzu (Pueraria lobota), in preventing these errors. Analyses will be made in embryonic, larval and adult stages of the animal. We will apply histological, histochemical and molecular biological techniques including quantitative RT-PCR and DNA methylation analysis for this study. This study will provide a foundation for identifying an alcohol metabolizing enzyme gene that is primarily responsible for FAS, and determine if a natural product will rescue the embryo from epigenetic ethanol toxicity. In this proposal, we have focused on an ethanol-induced epigenetic event as the major cause of Fetal Alcohol Syndrome and evaluate a natural product as the preventive agent for FAS.

描述（由申请人提供）：本文所述的项目的主要重点是研究乙醇对日本Medaka胚胎发生中基因表达的醛脱氢酶1A2（ALDH1A2）的影响，并确定该基因启动子促进剂区域中CPG岛甲基化模式的改变是否可能导致乙醇促成的乙醇杂种。乙醇在怀孕期间摄取酒精的母亲的新生婴儿中诱导胎儿酒精谱系障碍（FASD）。在FASD中，胎儿酒精综合征（FAS）是最临床上可识别的形式，该形式通过延迟，中枢神经系统（CNS）畸形和功能障碍，并且具有颅面，心血管疾病和肢体缺陷的独特模式。乙醇源自胎儿发育的分子机制尚不清楚。此外，除了在怀孕期间戒酒以外的女性以外，防止FAS的预防尚不清楚。以前，我们已经证明，在胚胎发生过程中暴露于乙醇的Medaka胚胎在颅面，心血管和骨骼器官中发展了早熟的FAS特征，这些特征可以与人类观察到的FAS特征进行比较。这些发现促使我们将Medaka用作搜索负责FA的基因的独特模型。我们的假设是酶醇脱氢酶（ADH）和醛脱氢酶（ALDH）的乙醇代谢会改变细胞NAD + /NADH比，并诱导氧化应激。这种改变的氧化还原态能够诱导特定基因的DNA分子的表观遗传修饰，从而调节胚胎的正常发育。在此应用中，我们将研究ALDH1A2基因启动子区域中CPG岛中的DNA甲基化模式，我们已将其确定为Medaka胚胎发生中的乙醇靶基因。我们还将确定天然产品Kudzu（pueraria lobota）在防止这些错误方面的影响。分析将在动物的胚胎，幼虫和成年阶段进行。我们将在本研究中应用组织学，组织化学和分子生物学技术，包括定量RT-PCR和DNA甲基化分析。这项研究将为识别主要负责FA的酒精代谢酶基因提供基础，并确定天然产物是否会从表观遗传乙醇毒性中挽救胚胎。在此提案中，我们将重点放在乙醇引起的表观遗传事件上，是胎儿酒精综合征的主要原因，并评估天然产物作为FAS的预防剂。