Employing subcellular calcium to control membrane voltage

利用亚细胞钙来控制膜电压

• 批准号: 9136155
• 负责人: UTE H HOCHGESCHWENDER
• 金额: $ 23.03万
• 依托单位: CENTRAL MICHIGAN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-02 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9136155
• 关键词: Address; Animals; Attenuated; Biochemical Reaction; Biological; Brain; Calcium; Calcium Binding; Calcium Channel; Cell membrane; Cells; Coupling; Detection; Development; Elements; Engineering; Enzymes; Event; Excision; Feasibility Studies; Feedback; Goals; Health; Human; Implant; Individual; Injection of therapeutic agent; Ion Pumps; Ions; Lead; Light; Link; Location; Luciferases; Membrane; Mental disorders; Methods; Mission; Molecular; Neurons; Opsin; Outcome; Peptides; Peripheral; Point Mutation; Production; Proteins; Proton Pump; Public Health; Research; Runaway; Self-control as a personality trait; Series; Source; Speed; Staging; System; Therapeutic; Time; Translating; biological systems; calcium indicator; clinically relevant; coelenterazine; design; improved; innovation; light emission; luciferin; nervous system disorder; neuroregulation; new technology; non-invasive imaging; novel; novel strategies; optogenetics; reconstitution; relating to nervous system; research study; sensor; tool; voltage

 DESCRIPTION (provided by applicant): The goal of this proposal is to assess the feasibility of an all-molecular method for activity-dependent feedback control of neuronal activity. We propose to generate calcium sensitive light emitting molecules (bioluminescent enzymes, luciferases) that drive light sensing optogenetic elements (channels or ion pumps, opsins) to control membrane voltage at the level of single cells for positive and negative feedback control. By adjusting calcium sensitivity and molecule location, light production can be made specific to large events such as bursts, or sensitive to individual spikes or single channel activity. By coupling these new luciferases to opsins, highly specific sensing of calcium at its source will trigger opsin activation for augmenting or suppressing neuronal activity, allowing a high degree of temporal and spatial precision in feedback control. Goals will be achieved by pursuing three aims: 1) Developing a calcium sensing split luciferase with significantly improved speed, brightness and range of sensitivity; 2) Targeting these new molecules to subcellular domains to enable highly specific biological outcomes; 3) Linking these new innovations to optogenetic readouts. Our strategy is non- invasive and it could be applied to large-scale manipulation of neural activity in behaving animals or in humans, where non-invasive, rapid feedback control of neuronal activity could be used to regulate clinically relevant activity in the brain. Our experiments are early stage, require proof of principle feasibility studies, but they have the potential to lead to a novel strategy to regulate activity only during periods of abnormal neuronal firing, such as attenuating runaway activity or amplifying local fluctuations. The molecular tools generated towards these feasibility experiments will be highly valuable in their own right, and achieving the goal of neural activity regulated self-control of neurons will be transformative.

 描述（由申请人提供）：该提案的目标是评估用于神经元活动的活动依赖性反馈控制的全分子方法的可行性。我们建议生成钙敏感的发光分子（生物发光酶、荧光素酶），驱动光传感光遗传元件（通道或离子泵、视蛋白）以控制单细胞水平的膜电压，以进行正反馈和负反馈控制。通过调整钙敏感性和分子位置，可以使光产生特定于爆发等大事件，或对单个尖峰或单通道活动敏感。通过将这些新的荧光素酶与视蛋白偶联，对钙源的高度特异性传感将触发视蛋白激活，从而增强或抑制神经元活动，从而在反馈控制中实现高度的时间和空间精度。目标将通过追求三个目标来实现：1）开发一种钙传感裂解荧光素酶，其速度、亮度和灵敏度范围显着提高； 2）将这些新分子靶向亚细胞域，以实现高度特异性的生物学结果； 3）将这些新创新与光遗传学读数联系起来。我们的策略是非侵入性的，它可以应用于行为动物或人类神经活动的大规模操纵，其中神经元活动的非侵入性、快速反馈控制可以用于调节大脑中的临床相关活动。我们的实验处于早期阶段，需要原理可行性研究证明，但它们有可能产生一种新策略，仅在神经元异常期间调节活动 发射，例如减弱失控活动或放大局部波动。这些可行性实验产生的分子工具本身就具有很高的价值，并且实现神经活动调节神经元自我控制的目标将是变革性的。