Advanced Dynamic Clamp for Neuroscience

用于神经科学的先进动态钳

• 批准号: 10213208
• 负责人: Mark W Nowak
• 金额: $ 5.5万
• 依托单位: CYTOCYBERNETICS, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-19 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10213208
• 关键词: Action Potentials; Address; Basic Science; Behavior; Binding; Calcium; Calcium-Activated Potassium Channel; Cells; Closure by clamp; Complex; Computer software; Data; Development; Disease; Electrical Engineering; Electronics; Electrophysiology (science); Engineering; Environment; Goals; Hodgkin-Huxley model; Home environment; Homeostasis; Injections; Interruption; Interview; Libraries; Ligands; Markov Chains; Membrane Potentials; Methodology; Methods; Modeling; Monitor; Morphologic artifacts; Muscle Cells; Myocardium; Neurons; Neurosciences; Operating System; Outcome; Patients; Pharmaceutical Preparations; Play; Potassium Channel; Production; Public Health; Quality Control; Recurrence; Reporting; Research; Research Personnel; Second Messenger Systems; Signal Transduction; Source; Speed; System; Technical Expertise; Technology Transfer; Testing; Time; Work; analog; base; cell type; commercial application; design; digital; drug discovery; experimental study; human tissue; induced pluripotent stem cell; innovation; markov model; mathematical model; pre-clinical; prevent; research and development; response; screening; simulation; skills; software systems; stem cells; user-friendly; voltage; voltage gated channel

The goal of this proposal is to develop a commercial “plug and play”, user-friendly, powerful and reliable dynamic clamp system, to enable all neuronal electrophysiologists to be able to perform sophisticated dynamic clamp experiments, without any requirement for programming, engineering, or mathematical modeling skills. We will develop hardware and software specifically for neuroscience applications, focusing on the specific stability and reliability needed for routine neuronal electrophysiology. We will focus on the critical combination of software, operating system, and hardware to achieve high speeds, but more importantly high reliability. Problems with timing, interrupts, and stability, were the key concerns and recurrent problems raised in interviews with our potential customers who were using home-made research platforms. Innovations in this proposal include: 1) A digital modulated conductance clamp mode and supporting software that will greatly expand the stability of the system during the rapid voltage changes that occur during neuronal action potentials; 2) The first dynamic clamp system to incorporate real-time Ca2+ transient (and other signaling) as an input. Many currents and related electrophysiological behaviors are sensitive to global Ca2+ levels. We will have the first dynamic clamp system capable of using Ca2+ (and other fluorometric signals) to interact with current amplitudes and gating behavior. This is a major advance in the application of dynamic clamp. 3) This first Markov Model based dynamic clamp system. Many channels are much more accurately modeled using Markov models as opposed to using the older and simpler Hodgkin-Huxley formalism. This is of critical importance in modeling ligand gated binding, Ca2+- dependent behavior and state-dependent drug binding. The three aims of this project are to 1) Develop rectifying background currents for the stabilization of neuronal resting potentials. We will establish a library of rectification models and a spline-driven model derived from user input that can be used in real-time during experiments. 2) Develop a Dynamic Clamp system with two inputs, the standard one for voltage, and an additional input for real time calcium transients in order to simulate channels such as calcium activated potassium channels. 3) Develop digitally controlled conductance clamp for enhanced stability. This aim will automatically prevent, detect, and correct data artefacts arising from the limitations of dynamic clamp methodology, which increases quality control.

本提案的目标是开发一种商业化的“即插即用”、用户友好、功能强大且可靠的动态 钳系统，使所有神经元电生理学家能够进行复杂的动态钳 实验，而无需任何编程，工程或数学建模技能的要求。我们将 开发专门用于神经科学应用的硬件和软件，重点关注特定的稳定性和 常规神经电生理学所需的可靠性。我们将专注于软件的关键组合， 操作系统和硬件实现高速度，但更重要的是高可靠性。问题 时间、中断和稳定性是我们在采访中提出的关键问题和经常性问题。 使用自制研究平台的潜在客户。本提案的创新之处包括：1）A 数字调制电导钳位模式及配套软件，将大大扩展 系统在神经元动作电位期间发生的快速电压变化期间; 2）第一动态钳位 系统将实时Ca 2+瞬变（和其他信号）作为输入。许多电流和相关 电生理行为对整体Ca 2+水平敏感。我们将有第一个动态夹紧系统 能够使用Ca 2+（和其他荧光信号）与电流幅度和门控行为相互作用。 这是动态夹具应用的一个重大进步。3)第一个基于马尔可夫模型的动态钳位 系统许多渠道更准确地建模使用马尔可夫模型，而不是使用旧的 更简单的Hodgkin-Huxley形式主义。这在模拟配体门控结合、Ca 2 +- 依赖行为和状态依赖药物结合。该项目的三个目标是 1)开发用于稳定神经元静息电位的整流背景电流。我们将建立一个 校正模型库和从用户输入导出的可实时使用的样条驱动模型 在实验中。 2)开发具有两个输入的动态箝位系统，标准输入用于电压，另一个输入用于 真实的时间钙瞬变以便模拟通道，例如钙激活钾通道。 3)开发数字控制的电导钳，以增强稳定性。这一目标将自动防止， 检测并纠正因动态钳位方法的局限性而产生的数据伪影，这会增加 质量控制