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非常重要。阿司匹林和其他非甾体抗炎药（NSAID）特异性抑制前列腺素合成，并与癌症预防和可逆性不孕症有关。前列腺素是排卵和受精所必需的，但其分子机制还不清楚。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。本申请的目的是确定线虫模型秀丽隐杆线虫可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，我们将测试的核心假设，即卵母细胞合成三尖杉酯碱，以调节排卵和精子的指导。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（PUFA），和预测的前列腺素脱氢酶是这些过程所必需的。此外，在不存在PUFA的情况下，野牡丹素可以调节精子活力。我们的第一个目标是描绘一个前列腺素生物合成途径，在卵母细胞中发挥作用。为了实现这一目标，我们将在遗传和生物化学水平上表征预测的前列腺素生物合成酶。我们的第二个目的是确定在何种程度上，野牡丹素和非甾体类抗炎药影响受精。为了实现这一目标，我们将研究在体内的精子活力和排卵的影响，三尖杉酯碱和NSAID。我们的最终目的是确定卵母细胞合成的PUFA衍生信号。遗传学和液相色谱法结合电喷雾串联质谱法将用于实现这一目标。我们期望，完成拟议的目标将建立C。作为第一个简单的，遗传学上易于处理的模型，发现生殖过程中前列腺素作用的分子机制。这些研究应该导致前列腺素合成和功能所需的基因的全面描绘，其中任何一个都可以编码新的药物靶点。前列腺素缺乏小鼠支持这一观点，即在哺乳动物中，前列腺素在受精中的作用可能是保守的。无论如何，对C。线虫的生殖腺具有发现人类不育、避孕和癌症的分子机制的潜力。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。对卵巢中的野牡丹素的基础研究有可能发现癌症、不育和避孕的分子机制。本建议的目的是确定在何种程度上的模式系统秀丽隐杆线虫可以用来阐明生殖过程中的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.