Laser Diode for Ion Channel Stimulation

用于离子通道刺激的激光二极管

• 批准号: 7555866
• 负责人: Mikhail I. Nemenov
• 金额: $ 10万
• 依托单位: LASMED, LLC
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-15 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7555866
• 关键词: Action Potentials; Analgesics; Asses; Binding; C Fiber; Calcium; Calibration; Capsaicin; Cells; Certification; Class; Computer software; Cultured Cells; Data; Development; Devices; Diagnosis; Electronics; Evaluation; Eye; Family; Fiber; Funding; Gated Ion Channel; Goals; Heating; Human; Image; In Vitro; Ion Channel; Lasers; Magnetism; Measurement; Measures; Mediating; Microscope; Modeling; Monitor; Nerve; Nerve Fibers; Nerve Tissue; Neurons; Nociceptors; Pain; Patients; Phase; Physiologic pulse; Process; Protein Family; Proteins; Protocols documentation; Pulse taking; Rate; Rattus; Ruthenium Red; Shapes; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Standards of Weights and Measures; TRPV1 gene; Temperature; Testing; United States Food and Drug Administration; capsazepine; concept; desensitization; design; drug discovery; in vivo; patch clamp; prototype; receptor; research study; tool; Action Potentials; Analgesics; Asses; Binding; C Fiber; Calcium; Calibration; Capsaicin; Cells; Certification; Class; Computer software; Cultured Cells; Data; Development; Devices; Diagnosis; Electronics; Evaluation; Eye; Family; Fiber; Funding; Gated Ion Channel; Goals; Heating; Human; Image; In Vitro; Ion Channel; Lasers; Magnetism; Measurement; Measures; Mediating; Microscope; Modeling; Monitor; Nerve; Nerve Fibers; Nerve Tissue; Neurons; Nociceptors; Pain; Patients; Phase; Physiologic pulse; Process; Protein Family; Proteins; Protocols documentation; Pulse taking; Rate; Rattus; Ruthenium Red; Shapes; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Standards of Weights and Measures; TRPV1 gene; Temperature; Testing; United States Food and Drug Administration; capsazepine; concept; desensitization; design; drug discovery; in vivo; patch clamp; prototype; receptor; research study; tool

The goal of this Phase II SBIR project is to develop and validate a laser diode stimulator to assess heat-sensitive Transient Receptor Potential (TRP) ion channels for in vitro and in vivo studies, using patch clamp recording and calcium imaging and compound action potential recording, respectively. This laser will be very useful for new drug discovery and evaluation, as there is currently no commercially available tool of similar capabilities. Evaluation of TRP family proteins can be useful in describing the potential utility of putative analgesic drugs. The ability to selectively (by temperature thresholds), reliably and reproducibly activate TRPs may also provide unique utility in understanding the mechanisms of antagonist action (binding) and provide additional information about mechanisms of interaction of different TRP proteins in cells and nerve tissue. There is also a potential utility of being able to differentially evaluate pain mediated by TRPV1 or TRPV2 proteins in the diagnosis and monitoring of progression of pain patients. Thus, it is our overall goal to establish stimulation protocols for use with a laser stimulator for in vitro and in vivo activation of heat gated ion channels. Laser stimulation is used for selective activation of C and A delta fibers in humans and rats (in our parallel projects), allowing reproducible protocols to test putative analgesic, from cell culture to humans. Thus, we hypothesize that the laser ion channel stimulator will prove safe and reliable action and will meet the FDA requirements for a laser device. To do this, we will develop an additional electronic circuit that allows the blockade of laser emission through the microscope, protecting eyes from any damage. We will also submit the laser and its components for certification of electrical and magnetic interference standards, and make any device changes that are necessary to meet the requirements. In addition, we will develop a protocol of direct temperature measurement of laser-induced temperature and modify the device to allow a tunable rate of heating. We will also insure the compatibility of the laser with calcium imaging and will continue to test the laser with TRPV1 DRG and HEK293 cells in order to study sensitization/desensitization processes. To do this we will apply brief and prolonged laser pulses and capsazepine. We will test the concept of TRP-associated activation of C and A delta fibers by direct laser activation of nerve fibers and application of the TRPV1 and TRPV2 antagonists capsazepine and ruthenium red.

该II阶段SBIR项目的目标是开发和验证激光二极管刺激器以评估 热敏感的瞬态受体电位（TRP）离子通道用于体外和体内研究，使用使用 斑块夹记录和钙成像和复合动作电势记录。 该激光器对于新药物发现和评估非常有用，因为目前没有 具有类似功能的市售工具。 TRP家族蛋白的评估可用于描述推定镇痛的潜在效用 毒品。可靠和可重复激活TRP的（按温度阈值）选择性（按温度阈值）的能力 还可以提供独特的效用，以理解拮抗剂作用的机制（结合）和 提供有关细胞中不同TRP蛋白相互作用机制的其他信息 神经组织。也有能力差异评估疼痛介导的疼痛的潜力 诊断和监测疼痛患者进展的TRPV1或TRPV2蛋白。因此，是 我们建立与激光刺激器一起用于体外和体内使用激光刺激器的刺激方案的总体目标 热门离子通道的激活。激光刺激用于选择性激活C和A 人类和大鼠的三角洲纤维（在我们的平行项目中），允许可重现协议测试 假定的镇痛药，从细胞培养到人类。 因此，我们假设激光离子通道刺激器将被证明是安全可靠的作用，并且将 满足FDA对激光设备的要求。为此，我们将开发额外的电子 可以通过显微镜阻断激光发射的电路，从而保护眼睛免受任何 损害。我们还将提交激光及其组件以证明电气和磁性 干扰标准，并进行满足要求所需的任何设备更改。 此外，我们将制定激光诱导的直接温度测量方案 温度并修改设备以使其加热速率。我们还将确保 激光与钙成像的兼容性，并将继续用TRPV1 DRG测试激光器 和HEK293细胞以研究敏化/脱敏过程。为此，我们将申请 简短和长时间的激光脉冲和辣椒粉。我们将测试与TRP相关的概念 通过直接激活神经纤维激活C和A Delta纤维的激活和应用 TRPV1和TRPV2拮抗剂辣椒粉和红色。