Ethanol effects on retinoic acid function in embryo hippocampus

乙醇对胚胎海马视黄酸功能的影响

• 批准号: 8006507
• 负责人: JOSEPH L NAPOLI
• 金额: $ 34万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-20 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8006507
• 关键词: Adult; Affect; Alcohol consumption; Alcohol dehydrogenase; Alcohols; All-Trans-Retinol; Area; Biological; Biological Assay; Blood alcohol level measurement; Brain; Brain region; Calories; Cerebellum; Chronic; Cognitive; Development; Embryo; Embryonic Development; Ethanol; Ethanol toxicity; Fetal Alcohol Syndrome; Gene Expression; Genetic Transcription; Genomics; Growth; Hippocampus (Brain); Hormonal; Intake; Knockout Mice; Learning; Life; Liver; Metabolism; Mus; Neuraxis; Neurons; Outcome; Pathology; Pathway interactions; Physiological; Process; Regulation; Retinoids; Sampling; Short-Term Memory; Signal Transduction; Staging; Testing; Testis; Tissues; Translations; Tretinoin; Use of New Techniques; Vitamin A; Work; alcohol effect; alcohol exposure; alcohol response; chronic alcohol ingestion; feeding; in vivo; insight; liquid chromatography mass spectrometry; nervous system development; neurogenesis; problem drinker

DESCRIPTION (provided by applicant): This project aims to determine the effects of chronic ethanol ingestion by dams on the concentration and function of atRA (all-trans-retinoic acid) in the embryo hippocampus during normal vitamin A nutriture, using new techniques for quantifying atRA and new insights into retinoid function in the hippocampus. The effects of ethanol on retinoid metabolism and function are poorly understood. The notion persists that ethanol inhibits bioactivation of retinol into atRA, but has not been tested directly in vivo during normal vitamin A nutriture. To test this premise, we developed an LC/MS/MS assay capable of quantifying atRA in specific brain areas. We determined during normal vitamin A nutriture that chronic ethanol exposure (36% of calories for 1 month) increases atRA in the adult mouse cortex (2-fold) and hippocampus (20-fold), without affecting other brain areas. Chronic ethanol exposure also increases atRA in the testis, but does not change atRA in several other mouse retinoid-target tissues. Chronic ethanol fed to dams starting e13 increases embryo atRA in the cortex (up to 50-fold) and the hippocampus (up to 20-fold), depending on blood alcohol content, quantified on e19. Emerging insight indicates that atRA affects learning and cognitive ability, besides nervous system development and neuron specification. Because excessive atRA produces teratogenic CNS effects, and the hippocampus relies on retinoid signaling to establish short-term memory, super physiological concentrations of atRA could contribute to ethanol toxicity, including fetal alcohol syndrome. We will test the hypothesis that ethanol-induced increases in embryo hippocampus atRA contributes to ethanol toxicity. Each aim will determine the effects of dam ethanol ingestion on embryo hippocampus retinoid concentrations and/or function. Wild type and CrbpI-null mice will be evaluated, because ethanol does not increase atRA in hippocampus of CrbpI-null mice. Aim 1 will determine retinoid concentrations in the embryo hippocampus during development. We will focus on e13 to e18 because hippocampus begins developing between e13-e15 and concludes in the CA1 region by e18. Aim 2 will test whether dam ethanol ingestion alters embryo atRA-regulated gene expression (transcription and translation). Aim 3 will test whether ethanol and/or increased atRA affect neurogenesis in the developing hippocampus. Aim 4 will test chronic ethanol effects on atRA-induced dendritic growth. We propose to answer: 1) what are the effects of chronic ethanol ingestion by dams on concentrations of atRA in the embryo hippocampus during normal vitamin A nutriture; 2) do ethanol- induced changes in atRA affect atRA-regulated transcription and translation, atRA processes as a whole (pathway and principle component analysis), or atRA-stimulated dendritic growth or neurogenesis? Even if the outcome reveals the impact of ethanol on atRA is not a major mechanism of its pathology, these studies will provide new insight into mechanisms of ethanol toxicity and retinoid function, and should influence approaches to treating alcoholics. PUBLIC HEALTH RELEVANCE: Vitamin A is essential for vertebrate life, because it is necessary for embryonic development, regulation of metabolism and function of the central nervous system, among many other functions. Alcohol (ethanol) ingestion causes severe depletion of vitamin A storage in the liver, and has been postulated to inhibit activation of vitamin A into its hormonal form, all-trans-retinoic acid (atRA). This project seeks to understand how chronic ethanol ingestion affects the concentrations of atRA in tissues during normal dietary vitamin A intake in the hippocampus area of the brain (the area needed to develop short-term memory), and to determine the effects of ethanol on atRA function in the hippocapus.

描述（由申请人提供）：本项目旨在确定母鼠长期摄入乙醇对正常维生素A营养期间胚胎海马中atRA（全反式维甲酸）浓度和功能的影响，使用新技术定量atRA和对海马中维甲酸功能的新见解。乙醇对类维生素A代谢和功能的影响知之甚少。这一观点坚持认为乙醇抑制视黄醇生物活化为atRA，但在正常维生素A营养期间尚未直接在体内进行测试。为了测试这一前提，我们开发了一种LC/MS/MS测定，能够定量特定脑区的atRA。我们确定在正常的维生素A营养期间，慢性乙醇暴露（1个月36%的卡路里）增加成年小鼠皮质（2倍）和海马（20倍）中的atRA，而不影响其他大脑区域。慢性乙醇暴露也增加了睾丸中的atRA，但不会改变其他几个小鼠类维生素A靶组织中的atRA。从e13开始向母鼠喂食慢性乙醇会增加皮质（高达50倍）和海马（高达20倍）中的胚胎atRA，这取决于在e19上定量的血液酒精含量。新的见解表明，atRA影响学习和认知能力，除了神经系统发育和神经元规格。由于过量的atRA产生致畸性CNS效应，并且海马依赖于类维生素A信号传导来建立短期记忆，因此超生理浓度的atRA可能导致乙醇毒性，包括胎儿酒精综合征。我们将测试乙醇诱导的胚胎海马atRA增加有助于乙醇毒性的假设。每个目标将确定母鼠乙醇摄入对胚胎海马类维生素A浓度和/或功能的影响。将评价野生型和CrbpI缺失小鼠，因为乙醇不会增加CrbpI缺失小鼠海马中的atRA。目的1将确定发育过程中胚胎海马中维甲酸的浓度。我们将重点放在e13到e18，因为海马体在e13-e15之间开始发育，并在e18结束于CA 1区域。目标2将测试母鼠乙醇摄入是否会改变胚胎atRA调节的基因表达（转录和翻译）。目标3将测试乙醇和/或增加的atRA是否影响发育中海马的神经发生。目的4将测试慢性乙醇对atRA诱导的树突状生长的影响。我们建议回答：1）在正常维生素A营养期间，母鼠长期摄入乙醇对胚胎海马中atRA浓度的影响是什么; 2）乙醇诱导的atRA变化是否影响atRA调节的转录和翻译、atRA过程作为一个整体（途径和主成分分析）或atRA刺激的树突生长或神经发生？即使结果显示乙醇对atRA的影响不是其病理学的主要机制，这些研究将为乙醇毒性和类维生素A功能的机制提供新的见解，并应影响治疗酗酒者的方法。 公共卫生关系：维生素A是脊椎动物生命所必需的，因为它是胚胎发育、代谢调节和中枢神经系统功能所必需的。酒精（乙醇）摄入会导致肝脏中维生素A储存的严重消耗，并被认为会抑制维生素A活化成其激素形式，全反式视黄酸（atRA）。该项目旨在了解慢性乙醇摄入如何影响正常饮食中维生素A摄入期间大脑海马区（开发短期记忆所需的区域）组织中atRA的浓度，并确定乙醇对atRA功能的影响。