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

摘要产前酒精暴露可通过凋亡机制诱导胎儿大脑神经元死亡。有证据表明，酒精暴露可能通过直接激活线粒体凋亡信号通路加速发育中的大脑细胞凋亡。尽管研究表明，产前酒精暴露会诱发线粒体功能障碍，但对酒精暴露对线粒体信号通路的影响知之甚少。事实上，大多数研究都集中在酒精暴露对线粒体解剖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、微阵列和其他分析来研究产前酒精暴露对线粒体信号通路的影响。此外，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包括酒精在内的氧化应激方面起着关键作用。一种是由活性依赖性神经营养因子（ADNF）衍生的9个氨基酸肽（salrsipa），称为SAL或ADNF-9；另一种是由活性依赖性神经保护蛋白（ADNP）衍生的8个氨基酸肽（NAPVSIPQ），称为NAP。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线粒体谷胱甘肽减少。在目前的建议中，我们将在胎儿酒精暴露（FAE）模型中使用我们建立的混合酒精的液体饮食，模拟适度饮酒。由于SAL和NAP保护线粒体免受酒精暴露的分子机制尚未完全确定，因此我们的目标是描绘精确的线粒体信号通路，介导胎儿大脑中SAL/NAP对产前酒精暴露的保护。基于体外和体内对线粒体氧化应激相关的研究，我们假设：1)产前酒精暴露通过涉及Bcl-2家族的线粒体信号通路诱导线粒体功能障碍并随后导致细胞死亡；2)产前酒精暴露改变细胞核和线粒体编码蛋白，随后诱导细胞凋亡；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。该应用程序产生的数据将详细了解产前酒精暴露对线粒体信号通路的影响，以及在FAE模型中ADNF-9和NAP在保护线粒体中的作用。

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.