Nitric Oxide and Responses to Hypoxia in Drosophila

一氧化氮和果蝇对缺氧的反应

• 批准号: 6771540
• 负责人: PATRICK H O'FARRELL
• 金额: $ 34.09万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6771540
• 关键词: Drosophilidae; RNA interference; arthropod genetics; genetic screening; high throughput technology; hypoxia; immune response; immunity; immunogenetics; nitric oxide

DESCRIPTION (provided by applicant): The discovery of nitric oxide as a regulator of blood pressure, which was awarded the Nobel prize in 1995, launched intense investigation into its function in human health. Recognizing the diversity, complexity and conservation of nitric oxide actions, studies in model organisms appear relevant and needed. In our studies of Drosophila, nitric oxide induced behavioral and physiological changes consistent with a conserved role in adaptation to low oxygen (hypoxia). Seeking other parallels to its function in mammals, we found that nitric oxide activates innate immune responses in Drosophila. We developed robust assays in which tagged transgenes report immune induction in larvae or cultured cells (S2 cells) in response to nitric oxide, or to bacteria, or to hypoxia. The responses of S2 cells can be blocked by inactivation of specific genes by RNA interference (RNAi). To exploit this powerful avenue for genetic dissection, we constructed a library of 7,200 RNAs representing the conserved genes of Drosophila. We propose high-throughput RNAi screens for genes contributing to immune induction. In preliminary work, we identified the genes involved in the response to bacterial components and will do the same for genes involved in the responses to nitric oxide and hypoxia. We will further exploit our assays to define the sequence of gene action, thereby delineating distinctions and commonalities in the pathways transducing these signals. Using in vivo genetics and tests in culture, we will place the signaling pathways in their biological context. Localization of function will position gene action in a cascade that conveys immune responses from the site of infection to distant tissues. These studies will provide new models for the action of signals central to human physiology and health.

描述（由申请人提供）：一氧化氮作为血压调节剂的发现，于1995年获得诺贝尔奖，启动了对其在人类健康中功能的深入研究。认识到多样性，复杂性和保护一氧化氮的行动，在模式生物的研究似乎相关和必要的。在我们对果蝇的研究中，一氧化氮诱导的行为和生理变化与适应低氧（缺氧）的保守作用一致。为了寻找与哺乳动物中的功能相似的其他功能，我们发现一氧化氮激活果蝇的先天免疫反应。我们开发了强大的检测，其中标记的转基因报告免疫诱导幼虫或培养细胞（S2细胞）响应一氧化氮，或细菌，或缺氧。S2细胞的反应可以通过RNA干扰（RNAi）使特定基因失活来阻断。为了利用这一强大的遗传解剖途径，我们构建了一个包含7,200个代表果蝇保守基因的RNA的文库。我们提出了高通量RNAi筛选有助于免疫诱导的基因。在初步工作中，我们确定了参与对细菌成分反应的基因，并将对参与对一氧化氮和缺氧反应的基因做同样的工作。我们将进一步利用我们的测定来确定基因作用的顺序，从而描绘出转导这些信号的途径的区别和共性。利用体内遗传学和培养测试，我们将把信号通路放在它们的生物学背景中。功能定位将基因作用定位在级联中，将免疫反应从感染部位传递到远处组织。这些研究将为人类生理和健康的核心信号的作用提供新的模型。