Prostaglandins in C. elegans Fertilization

描述(申请人提供)：前列腺素作用的分子机制对生殖生物学家、免疫学家、神经生物学家和肿瘤学家来说相当重要。阿司匹林和其他非类固醇抗炎药(NSAIDs)专门抑制前列腺素的合成，与癌症预防和可逆性不孕有关。前列腺素对排卵和受精是必不可少的，但其分子机制尚不清楚。小鼠是目前唯一可以用来研究前列腺素在生殖过程中功能的遗传系统。这项申请的目的是确定线虫模型秀丽线虫在多大程度上可以用来解决前列腺素生物学中的基本问题。为了实现这一目标，我们将检验中心假设，即卵母细胞合成前列腺素来调节排卵和精子引导。支持这一假设的初步数据显示，前列腺素前体、多不饱和脂肪酸(PUFAs)和预测的前列腺素合成酶是这些过程所必需的。此外，在没有多不饱和脂肪酸的情况下，前列腺素可以调节精子的活力。我们的第一个目标是描述在卵母细胞中发挥作用的前列腺素生物合成途径。为了实现这一目标，我们将在遗传和生化水平上对预测的前列腺素生物合成酶进行表征。我们的第二个目标是确定前列腺素和非甾体抗炎药对受精的影响程度。为了达到这一目的，我们将检测前列腺素和非甾体抗炎药对精子活力和体内排卵的影响。我们的最终目标是识别由卵母细胞合成的多不饱和脂肪酸衍生信号。为了实现这一目标，将使用遗传学和液相色谱与电喷雾电离串联质谱仪相结合的方法。我们预计，完成拟议的目标将使线虫性腺成为第一个简单的、遗传上容易处理的模型，用于发现生殖过程中前列腺素作用的分子机制。这些研究应该导致全面描述前列腺素合成和功能所需的基因，其中任何一个都可以编码新的药物靶点。前列腺素在受精中的作用在哺乳动物身上是可以保守的，这一观点得到了前列腺素缺陷小鼠的支持。无论如何，对线虫性腺中前列腺素的基础研究有可能发现人类不孕、避孕和癌症的分子机制。公共卫生相关性：前列腺素是对人类生殖、免疫和心血管功能至关重要的脂质信号。对卵巢前列腺素的基础研究有可能发现癌症、不孕症和避孕的分子机制。这项建议的目的是确定秀丽线虫模型系统在多大程度上可以用来阐明前列腺素在生殖过程中作用的分子机制。

DESCRIPTION (provided by applicant): The molecular mechanisms of prostaglandin action are of considerable importance to reproductive biologists, immunologists, neurobiologists, and oncologists. Aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs) specifically inhibit prostaglandin synthesis and are associated with cancer prevention and reversible infertility. Prostaglandins are essential for ovulation and fertilization, but the molecular mechanisms are not well understood. The mouse is the only genetic system currently available to study prostaglandin functions during reproduction. The objective of this application is to determine the extent to which the nematode model Caenorhabditis elegans can be used to address fundamental questions in prostaglandin biology. To achieve this objective, we will test the central hypothesis that oocytes synthesize prostaglandins to regulate ovulation and sperm guidance. Support for this hypothesis comes from preliminary data showing that prostaglandin precursors, polyunsaturated fatty acids (PUFAs), and predicted prostaglandin synthases are required for these processes. In addition, prostaglandins can regulate sperm motility in the absence of PUFAs. Our first aim is to delineate a prostaglandin biosynthesis pathway that functions in oocytes. To achieve this aim, we will characterize predicted prostaglandin biosynthesis enzymes at the genetic and biochemical levels. Our second aim is to determine the extent to which prostaglandins and NSAIDs influence fertilization. To achieve this aim, we will examine the effects of prostaglandins and NSAIDs on sperm motility and ovulation in vivo. Our final aim is to identify the PUFA-derived signals synthesized by oocytes. A combination of genetics and liquid chromatography coupled to electrospray ionization tandem mass spectrometry will be used to achieve this aim. We expect that completing the proposed aims will establish the C. elegans gonad as the first simple, genetically tractable model for discovering molecular mechanisms of prostaglandin action during reproduction. These studies should lead to a comprehensive delineation of genes that are required for prostaglandin synthesis and function, any of which could encode new drug targets. The role of prostaglandins in fertilization could be conserved in mammals, an idea supported by prostaglandin deficient mice. Regardless, basic research on prostaglandins in the C. elegans gonad has the potential for discovering molecular mechanisms of human infertility, contraception, and cancer. PUBLIC HEALTH RELEVANCE: Prostaglandins are lipid signals that are critical for human reproduction, immunity, and cardiovascular functions. Basic research on prostaglandins in the ovary has the potential for discovering molecular mechanisms of cancer, infertility, and contraception. The objective of this proposal is to determine the extent to which the model system Caenorhabditis elegans can be used to elucidate the molecular mechanisms of prostaglandin action during reproduction.