Calcium Channel Signaling in Neurons

神经元中的钙通道信号传导

• 批准号: 6757597
• 负责人: Ricardo E. Dolmetsch
• 金额: $ 29.27万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-15 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6757597
• 关键词: apoptosis; biological signal transduction; biophysics; cAMP response element binding protein; calcium channel; cell biology; cell differentiation; cell line; electrophysiology; gene expression; gene induction /repression; gene mutation; immunocytochemistry; immunoprecipitation; neural plasticity; neurobiology; neurons; polymerase chain reaction; posttranslational modifications; protein protein interaction; terminal nick end labeling; tissue /cell culture; transcription factor; voltage gated channel; western blottings; yeast two hybrid system

DESCRIPTION (provided by applicant): Voltage-gated calcium (Ca2+) channels play a central role in neuronal function and are essential for converting electrical activity into biochemical events. The main goal of this proposal is to identify the molecular mechanisms by which voltage-gated calcium channels activate signaling cascades that mediate gene expression and promote neuronal survival. Neurons and muscle cells express at least ten different kinds of voltage-gated calcium channels that vary in their subcellular localization and biophysical properties. L-type channels (LTCs) are particularly effective at activating transcriptional pathways and at promoting neuronal survival. The transcription factors CREB and MEF-2 are two important targets of LTC signaling that regulate differentiation and plasticity in the nervous system. The biophysical and biochemical features that allow LTCs to activate gene expression and suppress apoptosis are not well understood. To address the question of how LTCs are linked to signaling pathways the following specific aims are proposed: 1) To determine the structural and biophysical features that allow L-type calcium channels to activate transcription. 2) To determine whether specific L-type channel interacting proteins link the channel to signaling pathways that lead to the activation of transcription. 3) To determine what features of L-type calcium channels allow them to inhibit neuronal apoptosis and promote survival. Biochemical, cell biological and electrophysiological techniques will be used to develop these specific aims. We have recently developed a method of using dihydropyridine insensitive LTCs to investigate LTC signaling in primary neurons and we plan to use this approach for these studies. We have also identified several LTC-interacting proteins that may be important for channel regulation and signaling and we plan to investigate their importance for signaling to the nucleus. The results of these experiments will provide critical insights into how voltage-gated channels activate the signaling pathways that regulate the structure and function of the nervous system.

描述（由申请人提供）：电压门控钙 (Ca2+) 通道在神经元功能中发挥核心作用，对于将电活动转化为生化事件至关重要。该提案的主要目标是确定电压门控钙通道激活介导基因表达和促进神经元存活的信号级联的分子机制。神经元和肌肉细胞表达至少十种不同的电压门控钙通道，其亚细胞定位和生物物理特性各不相同。 L 型通道 (LTC) 在激活转录途径和促进神经元存活方面特别有效。转录因子 CREB ​​和 MEF-2 是 LTC 信号传导的两个重要靶标，调节神经系统的分化和可塑性。 LTC 激活基因表达和抑制细胞凋亡的生物物理和生化特征尚不清楚。为了解决 LTC 如何与信号通路联系的问题，提出了以下具体目标： 1) 确定允许 L 型钙通道激活转录的结构和生物物理特征。 2) 确定特定的L型通道相互作用蛋白是否将通道与导致转录激活的信号通路连接起来。 3) 确定L型钙通道的哪些特征使它们能够抑制神经元凋亡并促进存活。 将使用生物化学、细胞生物学和电生理学技术来实现这些具体目标。我们最近开发了一种使用二氢吡啶不敏感 LTC 来研究原代神经元中 LTC 信号传导的方法，我们计划使用这种方法进行这些研究。我们还鉴定了几种可能对通道调节和信号转导很重要的 LTC 相互作用蛋白，我们计划研究它们对细胞核信号转导的重要性。这些实验的结果将为电压门控通道如何激活调节神经系统结构和功能的信号通路提供重要的见解。