Regulation and Function of Cyclic GMP in the CNS

CNS 中环 GMP 的调节和功能

• 批准号: 6779704
• 负责人: DAVID B MORTON
• 金额: $ 30.2万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-18 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6779704
• 关键词: Caenorhabditis elegans; Drosophilidae; Manduca; antisense nucleic acid; arthropod genetics; central nervous system; cyclic GMP; ecdysone; enzyme activity; enzyme induction /repression; guanylate cyclase; hormone regulation /control mechanism; in situ hybridization; neurogenetics; nitric oxide; northern blottings; phosphoproteins; protein kinase; receptor; recombinant proteins; site directed mutagenesis

DESCRIPTION (provided by applicant): Almost all cellular functions are modulated by a variety of different intracellular messengers. This proposal focuses on the regulation and function of one of these messengers, cyclic GMP (cGMP). The balance of its synthesis and breakdown regulates the level of cGMP within a cell. The synthesis of cGMP is controlled by the enzyme guanylyl cyclase and until recently two main classes of guanylyl cyclase were known, receptor and soluble guanylyl cyclases. Receptor guanylyl cyclases are integral membrane proteins that are regulated by either extracellular peptide hormones or intracellular calcium-binding proteins. Guanylyl cyclases of this class regulate the cGMP levels in the kidney and in the photoreceptors of the retina. Soluble guanylyl cyclases are located in the cytoplasm and have classically been regulated by the gaseous messenger, nitric oxide. They are particularly important in regulating blood pressure via vascular smooth muscle tone. An example of the importance of cGMP in human health and disease is shown by the widespread use of the drug, sildenafil (Viagra) that elevates cGMP levels by inhibiting an enzyme that normally breaks down cGMP. Using an insect, Manduca sexta, as a model organism my lab has identified two novel classes of guanylyl cyclase. The first is related to receptor guanylyl cyclases but is not an integral membrane protein. The second is related to soluble guanylyl cyclases but is nitric oxide insensitive. We have evidence to suggest that this nitric oxide-insensitive soluble guanylyl cyclase mediates the action of a neuropeptide that regulates a specific behavioral program and associated physiological changes in Manduca and another insect, Drosophila melanogaster. The major focus of this proposal is to test this hypothesis directly combining the use of cellular and molecular approaches in Manduca with a genetic approach in Drosophila. In addition, we will use molecular and biochemical techniques to determine the biochemical cascade involved in the activation of this novel guanylyl cyclase. Members of these novel classes of guanylyl cyclase are also present in another model organism, Caenorhabditis elegans. Most signal transduction cascades are highly evolutionarily conserved and it is likely that examples of these guanylyl cyclases will be identified in mammals. A more detailed knowledge of the variety of mechanisms that regulate the levels of cGMP in different tissues will undoubtedly have a significant impact basic cellular physiology in a wide variety of systems.

描述（由申请人提供）： 几乎所有的细胞功能都受到多种不同的调节 细胞内信使。本提案重点关注监管和功能 这些信使之一，环鸟苷酸 (cGMP)。其合成与平衡 分解调节细胞内 cGMP 的水平。 cGMP的合成为 由鸟苷酸环化酶控制，直到最近有两个主要类别 已知鸟苷酸环化酶、受体和可溶性鸟苷酸环化酶。 受体鸟苷酸环化酶是整合膜蛋白，受以下因素调节 细胞外肽激素或细胞内钙结合 蛋白质。此类鸟苷酸环化酶调节 cGMP 水平 肾脏和视网膜的光感受器。可溶性鸟苷酸环化酶是 位于细胞质中，通常受气体调节 信使，一氧化氮。它们对于调节血液特别重要 通过血管平滑肌张力产生压力。 cGMP 重要性的一个例子 该药物的广泛使用表明了它对人类健康和疾病的影响， 西地那非（伟哥）通过抑制一种酶来提高 cGMP 水平 通常会分解 cGMP。 使用昆虫 Manduca sexta 作为模式生物，我的实验室已鉴定出两种 新类别的鸟苷酸环化酶。第一个与受体鸟苷基有关 环化酶，但不是完整的膜蛋白。第二个与 可溶性鸟苷酸环化酶，但对一氧化氮不敏感。我们有证据表明 表明这种一氧化氮不敏感的可溶性鸟苷酸环化酶介导 调节特定行为程序的神经肽的作用和 曼杜卡和另一种昆虫果蝇的相关生理变化 黑腹果蝇。该提案的主要重点是检验这一假设 直接将细胞和分子方法在 Manduca 中的使用与 果蝇的遗传方法。此外，我们将使用分子和 生化技术来确定参与的生化级联 这种新型鸟苷酸环化酶的激活。 这些新型鸟苷酸环化酶的成员也存在于另一种 模式生物，秀丽隐杆线虫。大多数信号转导级联是 进化上高度保守，这些例子很可能 将在哺乳动物中鉴定鸟苷酸环化酶。更详细的知识 不同组织中调节 cGMP 水平的多种机制 毫无疑问，基础细胞生理学将在广泛的领域产生重大影响。 各种系统。