Mechanism of Mechanoreceptor Inhibition by Aldosterone

醛固酮抑制机械感受器的机制

• 批准号: 6835702
• 负责人: Heather A Drummond
• 金额: $ 21.72万
• 依托单位: UNIVERSITY OF MISSISSIPPI MED CTR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6835702
• 关键词: RNA interference; afferent nerve; aldosterone; baroreceptors; baroreflex; biological signal transduction; calcium channel; genetic transcription; hormone regulation /control mechanism; laboratory mouse; mechanoreceptors; membrane channels; mitogen activated protein kinase; polymerase chain reaction; protein degradation; protein isoforms; receptor expression; sodium channel; tissue /cell culture

DESCRIPTION (provided by applicant): The long term objective of my lab is to understand the molecular basis of mechanotransduction. This includes the 1) identification of components of mechanosensitive ion channel complexes in sensory neurons and muscle cells, 2) regulation of mechanosensitive channel expression by physical and hormonal factors and 3) involvement in cardiovascular pathophysiology (autonomic dysfunction, hypertension). The current proposal will address a highly novel area of research: aldosterone regulation of mechanosensitive ion channel expression and function. Hypertension and chronic heart failure are characterized by elevations in circulating aldosterone. Aldosterone is known to inhibit baroreceptor function, which may contribute to cardiac arrhythmias frequently associated with these diseases. The molecular mechanism of aldosterone suppression of baroreflex control of the autonomic nervous system is unclear. The hypothesis underlying this proposal is that aldosterone activation of the MAPK signaling pathway, decreases the expression of mechanosensitive channels in baroreceptor neurons by inhibiting transcription and augmenting degradation of ENaC proteins, thus causing baroreceptor dysfunction. This is a novel hypothesis. The goals of this proposal are to determine if 1) aldosterone inhibits DEG/ENaC transcript expression and mechanically gated calcium transients, 2) activation of the MAPK signaling cascade inhibits ENaC expression and mechanically gated calcium transients, and 3) ENaC proteins are required for mechanosensory function in cultured sensory neurons. We will use real time RT-PCR and immunofluorescence to determine the role of aldosterone and MAPK pathway on ENaC expression in cultured sensory neurons. We will use 2 assays to assess mechanosensory function in cultured sensory neurons. The importance of ENaC proteins in mechanotransduction will be determined using interference RNA and overexpression of dominant-negative isoforms. The results from this proposal may help define the molecular mechanism of baroreceptor inhibition by aldosterone and provide a better understanding of the mechanism of autonomic dysfunction in hypertension and chronic heart failure.

描述(申请人提供)：我的实验室的长期目标是了解机械转导的分子基础。这包括1)识别感觉神经元和肌肉细胞中机械敏感离子通道复合体的组成，2)物理和激素因素对机械敏感通道表达的调节，3)参与心血管病理生理学(自主神经功能障碍，高血压)。目前的提案将涉及一个非常新的研究领域：醛固酮对机械敏感离子通道表达和功能的调节。高血压和慢性心力衰竭的特征是循环中的醛固酮升高。已知醛固酮抑制压力感受器功能，这可能导致经常与这些疾病相关的心律失常。醛固酮抑制自主神经系统压力感受性反射控制的分子机制尚不清楚。这一假设认为，醛固酮激活MAPK信号通路，通过抑制ENaC蛋白的转录和加速降解，减少压力感受器神经元机械敏感通道的表达，从而导致压力感受器功能障碍。这是一个新的假设。该方案的目的是确定1)醛固酮是否抑制DEG/ENaC转录本的表达和机械门控钙瞬变，2)MAPK信号通路的激活抑制ENaC的表达和机械门控钙瞬变，以及3)ENaC蛋白是培养的感觉神经元机械感觉功能所必需的。我们将使用实时定量RT-PCR和免疫荧光技术来确定醛固酮和MAPK通路对培养的感觉神经元ENaC表达的影响。我们将使用两种方法来评估培养的感觉神经元的机械感觉功能。ENaC蛋白在机械转导中的重要性将通过干扰RNA和显性-负性异构体的过度表达来确定。这一结果可能有助于确定醛固酮抑制压力感受器的分子机制，并为更好地理解高血压和慢性心力衰竭自主神经功能障碍的机制提供帮助。