The Role of Lipid Rafts in Fetal Alcohol Spectrum Disorder

脂筏在胎儿酒精谱系障碍中的作用

• 批准号: 7267884
• 负责人: CYNTHIA FRANCES BEARER
• 金额: $ 30.9万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7267884
• 关键词: Acids; Alcohol-related birth defects; Alcohols; Animals; Behavioral; Brain; Cell membrane; Cells; Cholesterol; Choline; Cysteine; Cytoplasmic Granules; Data; Defect; Development; Developmental Neurotoxicity of Ethanol; Dietary Intervention; Disruption; Endocytosis; Ethanol; Ethanol toxicity; Fetal Alcohol Spectrum Disorder; Fetal Alcohol Syndrome; Figs - dietary; Folic Acid; Ganglioside GM1; Growth Cones; Heavy Drinking; Homing; Human; In Vitro; Incidence; Infant; Intervention; Kinetics; Live Birth; Localized; Measures; Mediating; Membrane Microdomains; Mental Retardation; Modeling; Mus; NAPVSIPQ peptide; Nervous system structure; Neural Cell Adhesion Molecule L1; Neural Tube Defects; Neuraxis; Neurites; Neurons; Newborn Infant; Nutrient; Outcome; Pathway interactions; Peripheral; Pregnancy; Protective Agents; Proteins; Psychological Tests; Public Health; Rattus; Recycling; Research; Research Personnel; Role; Secondary to; Signal Transduction; Site; Supplementation; System; Teratogens; Test Result; Testing; Thinking; Toxic effect; Trefoil Motif; Work; alcohol effect; alcohol exposure; alcohol sensitivity; base; cost; drinking; human study; in vivo; in vivo Model; mutant; nervous system development; neurofascin; neurotoxicity; prevent; programs; protein distribution; protein protein interaction; pup; trafficking

DESCRIPTION (provided by applicant): Ethanol is a known human teratogen of immense public health impact. Heavy drinking during pregnancy is the leading known cause of mental retardation, while more subtle effects particularly on the developing brain occur in 1% of all liveborns. Recent evidence has shown that many of the known protein targets for ethanol's toxic effects on the developing central nervous system are associated with lipid rafts. Lipid rafts are specialized microdomains of the plasma membrane and serve as platforms for protein-protein interactions. Our prior work has shown that the ability of L1 cell adhesion molecule (L1), a protein critical for the proper development of the nervous system, to promote neurite outgrowth is exquisitely sensitive to ethanol. Localized disruption of lipid rafts reduces L1 mediated neurite outgrowth. Our preliminary data show that ethanol alters the distribution of L1 in lipid rafts, and that cholesterol depletion abolishes ethanol sensitivity. Further, our preliminary data suggests that other protein targets of ethanol change their lipid raft distribution in the presence of ethanol. We hypothesize that a major mechanism of ethanol toxicity is through alteration of lipid raft-protein interactions, and that interventions which reduce toxicity prevent this disruption. These hypotheses will be tested using cerebellar granule neurons (CGN) from rat pups and from mice, both wild type and lacking L1, and a rat in vivo entubation model. In Aim 1, we will confirm that lipid raft-L1 interactions underlie ethanol sensitivity of neurite outgrowth by transfecting CGN lacking L1 with mutant constructs of L1 lacking a signal to colocalize with lipid rafts. Neurite outgrowth in transfected cells is expected to be ethanol insensitive. In Aim 2, we will determine whether L1 or the lipid raft is the site of the ethanol action, and the effect of ethanol on the kinetics of L1 trafficking to and out of the lipid raft compartment. In Aim 3, we will use four nutrients shown to be protective in other experimental systems, to prevent the changes in protein distribution secondary to ethanol. The most effective combination of these nutrients will be determined with L1 in CGN and in the in vivo model. The relevance of this research is that simple dietary manipulations with these nutrients may prevent some aspects of ethanol neurotoxicity much like folic acid supplementation and reduction of neural tube defects.

描述（由申请方提供）：乙醇是一种已知的人类致畸剂，对公共卫生有巨大影响。怀孕期间大量饮酒是导致智力迟钝的主要已知原因，而1%的孕妇会出现更微妙的影响，特别是对发育中的大脑的影响。最近的证据表明，乙醇对发育中的中枢神经系统的毒性作用的许多已知蛋白质靶点与脂筏有关。脂筏是质膜上的特殊微区，是蛋白质相互作用的平台。我们先前的工作表明，L1细胞粘附分子（L1），一种对神经系统正常发育至关重要的蛋白质，促进神经突生长的能力对乙醇非常敏感。局部破坏脂筏减少L1介导的神经突生长。我们的初步数据表明，乙醇改变了L1在脂筏中的分布，胆固醇消耗废除乙醇敏感性。此外，我们的初步数据表明，乙醇的其他蛋白质目标改变其脂筏分布在乙醇的存在下。我们假设乙醇毒性的主要机制是通过改变脂筏-蛋白质相互作用，而降低毒性的干预措施可以防止这种破坏。将使用来自大鼠幼仔和小鼠（野生型和缺乏L1）的小脑颗粒神经元（CGN）以及大鼠体内插管模型来检验这些假设。在目标1中，我们将确认脂筏-L1相互作用的神经突生长的乙醇敏感性的基础，通过用缺乏与脂筏共定位的信号的L1的突变体构建体转染缺乏L1的CGN。预期转染细胞中的神经突生长是乙醇不敏感的。在目标2中，我们将确定L1或脂筏是否是乙醇作用的位点，以及乙醇对L1运输进出脂筏隔室的动力学的影响。在目标3中，我们将使用四种在其他实验系统中显示出保护作用的营养素，以防止乙醇引起的蛋白质分布变化。这些营养素的最有效组合将在CGN和体内模型中用L1确定。这项研究的相关性是，简单的饮食操作与这些营养素可能会防止某些方面的乙醇神经毒性，就像叶酸补充剂和减少神经管缺陷。