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）：该项目旨在确定大坝摄入慢性乙醇对正常维生素A营养期间胚胎海马中ATRA（全反击毒性酸）的浓度和功能的影响，该新技术使用新的技术来量化ATRA和新的洞察力，以量化Hipippopocampampus中的RetinIniof函数。乙醇对类维生素性代谢和功能的影响知之甚少。这个观念坚持认为，乙醇抑制视网膜生物活化到ATRA中，但在正常维生素A营养期间尚未直接在体内进行测试。为了测试这个前提，我们开发了一个能够在特定大脑区域量化ATRA的LC/MS/MS分析。我们确定在正常维生素A营养期间，慢性乙醇暴露（占1个月卡路里的36％）会增加成年小鼠皮层（2倍）和海马（20倍）的ATRA，而不会影响其他大脑区域。慢性乙醇暴露也会增加睾丸中的ATRA，但在其他几种小鼠类维生素类靶性组织中不会改变ATRA。喂入E13的慢性乙醇会增加皮质中的胚胎ATRA（最多50倍）和海马（最多20倍），具体取决于血液酒精含量，在E19上进行了定量。新兴的见解表明，除了神经系统发展和神经元规范外，ATRA会影响学习和认知能力。由于过度ATRA会产生致病性中枢神经系统作用，因此海马依赖于类维生素性信号来建立短期记忆，因此ATRA的超级生理浓度可能有助于乙醇毒性，包括胎儿酒精综合征。我们将检验以下假设：乙醇诱导的胚胎海马ATRA增加有助于乙醇毒性。每个目的都将确定大坝乙醇摄入对胚胎海马类维生素类动物浓度和/或功能的影响。将评估野生型和crbpi-null小鼠，因为乙醇不会增加CRBPI无效小鼠海马的ATRA。 AIM 1将确定发育过程中胚胎海马中的类视黄素浓度。我们将重点关注E13至E18，因为海马开始在E13-E15之间发展，并通过E18在CA1区域结束。 AIM 2将测试DAM乙醇摄入是否会改变胚胎ATRA调节的基因表达（转录和翻译）。 AIM 3将测试乙醇和/或ATRA是否会影响发展中海马的神经发生。 AIM 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，大坝摄入慢性乙醇对胚胎海马浓度的影响是什么？ 2）乙醇诱导的ATRA的变化是否会影响ATRA调节的转录和翻译，ATRA过程（途径和原理分析）或ATRA刺激的树突状生长或神经发生？即使结果揭示了乙醇对ATRA的影响并不是其病理的主要机制，这些研究也将提供对乙醇毒性和类视黄素功能机制的新见解，并应影响治疗酒精中毒的方法。 公共卫生相关性：维生素A对于脊椎动物的生活至关重要，因为这对于胚胎发展，中枢神经系统的新陈代谢和功能以及许多其他功能都是必要的。酒精（乙醇）摄入会导致肝脏中维生素A储存的严重消耗，并被认为是抑制维生素A激活其激素形式，即全反式反激酸（ATRA）。该项目旨在了解在正常饮食中的饮食维生素A摄入脑在大脑海马区域（发展短期记忆所需的区域），慢性乙醇摄入如何影响组织中ATRA的浓度，并确定乙醇对河马中ATRA功能的影响。