Prostaglandins in C. elegans Fertilization

描述（由申请人提供）：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具有相当重要的意义。阿司匹林和其他非甾体抗炎药（NSAIDs）特异性抑制前列腺素合成，与癌症预防和可逆性不孕症有关。前列腺素对排卵和受精至关重要，但其分子机制尚不清楚。小鼠是目前唯一可用于研究生殖过程中前列腺素功能的遗传系统。本应用程序的目的是确定线虫模型秀丽隐杆线虫可以用于解决前列腺素生物学中的基本问题的程度。为了实现这一目标，我们将验证卵母细胞合成前列腺素来调节排卵和精子引导的中心假设。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，前列腺素可以在缺乏PUFAs的情况下调节精子的活力。我们的第一个目标是描述在卵母细胞中起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表征预测的前列腺素生物合成酶。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。为了达到这个目的，我们将在体内研究前列腺素和非甾体抗炎药对精子运动和排卵的影响。我们的最终目的是鉴定卵母细胞合成的pufa衍生信号。遗传学和液相色谱结合电喷雾电离串联质谱法将用于实现这一目标。我们期望完成提出的目标将使秀丽隐杆线虫性腺成为第一个简单的，遗传上可处理的模型，用于发现前列腺素在生殖过程中的作用的分子机制。这些研究将导致对前列腺素合成和功能所需基因的全面描述，其中任何一个基因都可以编码新的药物靶点。前列腺素在哺乳动物受精中的作用可能是保守的，这一观点得到了前列腺素缺乏小鼠的支持。无论如何，对秀丽隐杆线虫生殖腺中前列腺素的基础研究有可能发现人类不育、避孕和癌症的分子机制。公共卫生相关性：前列腺素是一种脂质信号，对人类生殖、免疫和心血管功能至关重要。卵巢中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。本提案的目的是确定在何种程度上模型系统秀丽隐杆线虫可以用来阐明在繁殖过程中前列腺素作用的分子机制。

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.