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: 7414075
• 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 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7414075
• 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（由申请人提供）：乙醇是一种已知的人类致畸物，对公众健康有巨大影响。怀孕期间大量饮酒是已知的导致智力迟钝的主要原因，而在所有活产儿中，有1%的人会对发育中的大脑产生更微妙的影响。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都与脂筏有关。脂筏是质膜的特殊微结构域，是蛋白质相互作用的平台。我们之前的研究表明，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。脂筏的局部破坏减少L1介导的神经突生长。我们的初步数据表明，乙醇改变L1在脂筏中的分布，并且胆固醇消耗消除了乙醇敏感性。此外，我们的初步数据表明，乙醇的其他蛋白质靶点在乙醇存在下改变了它们的脂质筏分布。我们假设乙醇毒性的主要机制是通过脂筏-蛋白质相互作用的改变，而减少毒性的干预措施可以防止这种破坏。这些假设将使用来自大鼠幼崽和小鼠的小脑颗粒神经元（CGN）进行验证，包括野生型和缺乏L1的小鼠，以及大鼠体内插管模型。在Aim 1中，我们将通过转染缺乏L1的CGN和缺乏信号的L1突变体来与脂质筏共定位，证实脂质筏-L1相互作用是神经突生长的乙醇敏感性的基础。在转染的细胞中，神经突的生长预计对乙醇不敏感。在目标2中，我们将确定L1或脂质筏是乙醇作用的部位，以及乙醇对L1进出脂质筏室的运输动力学的影响。在目标3中，我们将使用在其他实验系统中显示具有保护作用的四种营养素，以防止继乙醇之后蛋白质分布的变化。这些营养素的最有效组合将在CGN和体内模型中通过L1来确定。这项研究的相关性在于，对这些营养物质进行简单的饮食控制，可能会预防乙醇神经毒性的某些方面，就像补充叶酸和减少神经管缺陷一样。