Molecular mechanism of ADNF and ADNP peptides in prevention of mitochondrial dysf

描述（由申请人提供）：摘要产前酒精暴露通过细胞凋亡机制诱导胎儿脑中的神经元死亡。有证据表明，酒精暴露可通过直接激活线粒体凋亡信号通路加速发育中大脑的凋亡。虽然研究表明，产前酒精暴露诱导线粒体功能障碍，相对较少的是知道酒精暴露对线粒体信号通路的影响。事实上，大多数研究都集中在酒精暴露对线粒体解剖结构变化和酶活性的影响上。相反，我们将使用最先进的蛋白质组学技术，微阵列和其他检测方法来研究产前酒精暴露对线粒体信号通路的影响。此外，在NIH-NICHD和特拉维夫大学进行的研究表明，两种肽在保护线粒体免受氧化应激（包括酒精）方面发挥关键作用。一种是九个氨基酸的肽（SALLRSIPA），称为SAL或ADNF-9，衍生自活性依赖性神经营养因子（ADNF）;另一种是八个氨基酸的肽（NAPVSIPQ），称为NAP，衍生自活性依赖性神经保护蛋白（ADNP）。在胎儿酒精综合征（FAS）模型中，这些肽已被证明可以防止酒精诱导的还原型线粒体谷胱甘肽减少。在目前的建议中，我们将使用我们建立的液体饮食与酒精混合的胎儿酒精暴露（FAE）模型，模仿适度饮酒。由于SAL和NAP保护线粒体免受酒精暴露的分子作用机制尚未完全确定，我们的目标是描绘精确的线粒体信号通路，介导SAL/NAP保护胎儿大脑免受产前酒精暴露。基于线粒体氧化应激的体内外研究，我们推测：1）产前酒精暴露通过Bcl-2家族的线粒体信号通路诱导线粒体功能障碍和细胞死亡：2）产前酒精暴露改变核和线粒体编码蛋白，诱导细胞凋亡; 3）ADNF-9和/或NAP直接和/或间接保护线粒体功能障碍免受产前酒精暴露的损害。本申请产生的数据将提供产前酒精暴露对线粒体信号通路的影响以及ADNF-9和NAP在FAE模型中保护线粒体的作用的详细了解。 公共卫生相关性：这项研究将调查怀孕期间酒精暴露对细胞中起呼吸作用的细胞器“线粒体”的影响。我们还将研究在怀孕阶段对中枢神经系统的“线粒体”有保护作用的分子对酒精暴露的作用。这将为寻找治疗胎儿酒精暴露的治疗方法提供信息。

DESCRIPTION (provided by applicant): Abstract Prenatal alcohol exposure induces neuronal death in the fetal brain through apoptotic mechanism. Evidence shows that alcohol exposure may accelerate apoptosis in the developing brain through direct activation of mitochondrial apoptotic signaling pathways. Although studies have demonstrated that prenatal alcohol exposure induces mitochondrial dysfunction, relatively little is known about the effects of alcohol exposure on mitochondrial signaling pathways. In fact, most studies have focused on the effects of alcohol exposure on mitochondrial anatomical changes and enzymes activities. In contrast, we will use state-of-the-art proteomics technology, microarray, and other assays to investigate the effects of prenatal alcohol exposure on mitochondrial signaling pathways. In addition, studies conducted at NIH-NICHD and Tel Aviv University have suggested that two peptides play a key role in protecting mitochondria against oxidative stress including alcohol. One is a nine amino acid peptide (SALLRSIPA), known as SAL or ADNF-9, derived from activity- dependent neurotrophic factor (ADNF); the other is an eight amino acid peptide (NAPVSIPQ), known as NAP, derived from activity-dependent neuroprotective protein (ADNP). In a model of fetal alcohol syndrome (FAS), these peptides have been shown to prevent alcohol-induced decreases in reduced mitochondrial glutathione. In the present proposal, we will use our established liquid diet mixed with alcohol in a model of fetal alcohol exposure (FAE) that mimics moderate alcohol drinking. Because the molecular mechanisms of action of both SAL and NAP in protecting mitochondria against alcohol exposure are not fully characterized, we aim to delineate the precise mitochondrial signaling pathways that mediate SAL/NAP protection against prenatal alcohol exposure in the fetal brain. Based on in vitro and in vivo studies on mitochondria related to oxidative stress, we hypothesize that: 1) Prenatal alcohol exposure induces mitochondrial dysfunction and subsequently cell death through mitochondrial signaling pathways involving Bcl-2 family; 2) Prenatal alcohol exposure alters nuclear and mitochondrial encoded proteins to subsequently induce apoptosis; 3) ADNF-9 and/or NAP protect directly and/or indirectly mitochondrial dysfunction against the insult of prenatal alcohol exposure. The data generated from this application will provide a detailed understanding of the effects of prenatal alcohol exposure on mitochondrial signaling pathways and the role of ADNF-9 and NAP in protecting mitochondria in a model of FAE. Public Health Relevance: This study will investigate the effect or alcohol exposure during pregnancy on an organelle termed "mitochondria' which play a respiratory role in cells. We will also investigate the role of molecules that have a protective effect against alcohol exposure on "mitochondria" of the central nervous system during pregnancy stages. This will provide information in finding therapeutic ways for the treatment of fetal alcohol exposure.