Advanced Dynamic Clamp for Neuroscience

用于神经科学的先进动态钳

基本信息

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
项目类别：
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

项目摘要

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.

该提案的目标是开发一种商业化的“即插即用”、用户友好、强大且可靠的动态钳系统，使所有神经电生理学家能够执行复杂的动态钳实验，无需任何编程、工程或数学建模技能。开发专门针对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重点关注软件的关键组合，操作系统和硬件要实现高速度，但更重要的是高可靠性的问题。时间、中断和稳定性是我们在采访中提出的关键问题和反复出现的问题。使用自制研究平台的潜在客户该提案的创新包括：1) A。数字调制电导钳模式及配套软件，将大大扩展稳定性 2）第一个动态钳位系统将实时 Ca2+ 瞬态（和其他信号）作为输入。电生理行为对整体 Ca2+ 水平敏感我们将拥有第一个动态钳系统。能够使用 Ca2+（和其他荧光信号）与电流幅度和门控行为相互作用。这是动态钳位应用中的一个重大进步 3) 这是第一个基于马尔可夫模型的动态钳位。与使用旧的系统相比，使用马尔可夫模型对许多通道进行建模要准确得多。和更简单的 Hodgkin-Huxley 形式主义对于模拟配体门控结合 Ca2+- 至关重要。该项目的三个目标是依赖行为和状态依赖药物结合。 1) 开发整流背景电流以稳定神经静息电位。整流模型库和从用户输入导出的可实时使用的样条驱动模型实验期间。 2) 开发一个具有两个输入的动态钳位系统，一个用于电压的标准输入，一个用于电压的附加输入实时钙瞬变，以模拟钙钾激活通道等通道。 3) 开发数控电导钳以增强稳定性这一目标将自动防止，检测并纠正由于动态钳方法的局限性而产生的数据伪影，这增加了质量控制。