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

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.

第二阶段 SBIR 项目的目标是开发和验证激光二极管刺激器以评估 用于体外和体内研究的热敏瞬时受体电位 (TRP) 离子通道，使用 分别是膜片钳记录和钙成像以及复合动作电位记录。 这种激光对于新药的发现和评估非常有用，因为目前还没有 具有类似功能的商用工具。 TRP 家族蛋白的评估可用于描述假定镇痛药的潜在效用 药物。能够选择性地（通过温度阈值）、可靠且可重复地激活 TRP 还可能为理解拮抗剂作用机制（结合）和 提供有关细胞中不同 TRP 蛋白相互作用机制的更多信息 神经组织。能够差异化评估由以下因素介导的疼痛也具有潜在效用： TRPV1 或 TRPV2 蛋白在疼痛患者的诊断和进展监测中的应用。因此，它是 我们的总体目标是建立与激光刺激器一起用于体外和体内的刺激方案 热门控离子通道的激活。激光刺激用于选择性激活 C 和 A 人类和大鼠的 delta 纤维（在我们的平行项目中），允许可重复的方案进行测试 假定的镇痛剂，从细胞培养物到人类。 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，并且将 满足 FDA 对激光设备的要求。为此，我们将开发一个额外的电子 允许阻挡通过显微镜的激光发射的电路，保护眼睛免受任何伤害 损害。我们还将提交激光器及其组件进行电磁认证 干扰标准，并进行任何必要的设备更改以满足要求。 此外，我们将开发激光诱导直接温度测量协议 温度并修改设备以允许可调的加热速率。我们还将确保 激光与钙成像的兼容性，并将继续使用 TRPV1 DRG 测试激光 和 HEK293 细胞，以研究致敏/脱敏过程。为此，我们将申请 短暂和长时间的激光脉冲和辣椒素。我们将测试 TRP 相关的概念 通过直接激光激活神经纤维来激活 C 和 A delta 纤维以及应用 TRPV1 和 TRPV2 拮抗剂辣椒西平和钌红。

项目成果

Sensory small fiber function differentially assessed with diode laser (DL) quantitative sensory testing (QST) in painful neuropathy (PN).

使用二极管激光 (DL) 定量感觉测试 (QST) 对疼痛性神经病 (PN) 的感觉小纤维功能进行差异评估。

• DOI: 10.1111/pme.12049
• 发表时间: 2013
• 期刊: Pain medicine (Malden, Mass.)
• 作者: Moeller-Bertram,Tobias;Schilling,JanM;Bačkonja,MiroslavMisha;Nemenov,MichaelI
• 通讯作者: Nemenov,MichaelI

The modality-specific contribution of peptidergic and non-peptidergic nociceptors is manifest at the level of dorsal horn nociresponsive neurons.

肽能和非肽能伤害感受器的模式特异性贡献在背角伤害反应神经元水平上表现出来。

• DOI: 10.1113/jphysiol.2012.242115
• 发表时间: 2013
• 期刊: The Journal of physiology
• 作者: Zhang,Jie;Cavanaugh,DanielJ;Nemenov,MichaelI;Basbaum,AllanI
• 通讯作者: Basbaum,AllanI