Force Clamp Systems for Evaluation of Mechanotransduction

用于评估机械传导的力夹系统

• 批准号: 7465346
• 负责人: Beth L Pruitt
• 金额: $ 30.06万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7465346
• 关键词: Accounting; Acquired Immunodeficiency Syndrome; Affect; Agar; Animals; Atomic Force Microscopy; Behavior; Behavioral; Biochemical; Biocompatible Materials; Biological; Biological Assay; Biological Models; Biomechanics; Caenorhabditis elegans; Cell membrane; Cell physiology; Cells; Communities; Complex; Coupling; Custom; Cytoskeleton; Data; Development; Devices; Diabetes Mellitus; Disease; Drug Design; Electronics; Electrophysiology (science); Esthesia; Evaluation; Exhibits; Extracellular Matrix; Feedback; Financial compensation; Fire - disasters; Floor; Freedom; Frequencies; Gated Ion Channel; Generations; Glass; Gold; Hair Cells; Health Care Costs; Image; In Vitro; Inherited; Ion Channel; Ions; Kinetics; Letters; Malignant Neoplasms; Measurement; Measures; Mechanics; Mediating; Methods; Modeling; Molecular; Mutation; Nematoda; Neurons; Neuropathy; Noise; Output; Pain; Pattern; Performance; Peripheral; Pharmaceutical Preparations; Pharmacotherapy; Physical Stimulation; Physiological; Physiology; Play; Process; Property; Proprioception; Public Health; Quantitative Evaluations; Ramp; Range; Rate; Reaction Time; Relaxation; Research; Research Personnel; Role; Scanning; Sensory; Shapes; Signal Transduction; Silicon; Skin; Spinal Ganglia; Standards of Weights and Measures; Stimulus; Structure; Surface; System; Techniques; Technology; Temperature; Testing; Therapeutic; Time; Tissues; Touch sensation; Travel; Variant; Vertebrates; Wood material; Work; base; cantilever; cost; design; desire; in vivo; innovation; insight; instrumentation; laser tweezer; microsystems; mutant; nano; novel; optical imaging; patch clamp; programs; prototype; receptor; reconstitution; research study; response; sensor; sensory neuropathy; somatosensory; tool; viscoelasticity; voltage

DESCRIPTION (provided by applicant): This work proposes a fully integrated approach to the study of mechano-electrical transduction in the sense of touch and focuses on the development of a novel microsystems-based tool-namely, a force clamp that uses appropriately-scaled piezeoresistive cantilevers as direct force sensors (PR force clamp). The proposal includes work to fully characterize the capabilities of a first-generation device, to design and build a second- generation device capable of operating at higher bandwidth, and to use these devices to analyze and model the biomechanics offeree transfer and mechano-electrical transduction by a model mechahoreceptor cell, the touch receptor neurons that innervate the body wall of the nematode Caenorhabditis elegans. C. elegans is an excellent biological platform for developing the proposed tools, which are likely to find application in the study of mechano-electrical transduction in other mechanosensory cells including vertebrate hair cells and dorsal root ganglion. A set of only six touch receptor neurons is responsible for behavioral responses to touch in C. elegans (compared to tens of thousands in vertebrates) and such responses are initiated by activation of the MEC-4 channel complex by forces applied to the body wall. Prototype force clamp systems use PR force clamps and piezoelectric actuators with programmable controllers to apply calibrated nano- to micro-Newton point load profiles with > 1kHz bandwidth. A critical and innovative aspect of this work is the integration of the PR force clamp with patch-clamp electrophysiology for synchronous force-displacement- physiological recordings. This technology will enable the first direct measurement of dynamic changes in tissue stiffness that may occur during mechano-electrical transduction and give rise to adaptation. Relevance to public health. The sense of touch can be degraded by both inherited and acquired disease, including AIDS and diabetes, as well as by chemotherapeutic drugs. Complications from such peripheral sensory neuropathies are estimated to cost more than $4 billion annual in health care costs. Despite this, the sense of touch and its degradation in disease remain poorly understood. The research tools developed in the course of this work have the potential to significantly advance our understanding.

描述（由申请人提供）：这项工作提出了一种完全集成的方法来研究触觉中的机电转换，并重点开发一种基于微系统的新型工具，即使用适当缩放的力夹压阻悬臂梁作为直接力传感器（PR 力夹）。该提案包括充分表征第一代设备的功能、设计和构建能够在更高带宽下运行的第二代设备以及使用这些设备对生物力学传递和机电转换进行分析和建模的工作模型机械感受器细胞是支配线虫体壁的触觉感受器神经元。线虫是开发所提出的工具的绝佳生物平台，这些工具很可能在其他机械感觉细胞（包括脊椎动物毛细胞和背根神经节）的机械电转导研究中得到应用。一组仅六个触觉感受器神经元负责线虫对触觉的行为反应（相比之下，脊椎动物中有数以万计），并且这种反应是通过施加到体壁的力激活 MEC-4 通道复合体来启动的。原型力夹系统使用 PR 力夹和带有可编程控制器的压电执行器，以应用带宽 > 1kHz 的校准纳米级到微米级牛顿点负载曲线。这项工作的一个关键和创新方面是将 PR 力钳与膜片钳电生理学相结合，以实现同步力-位移-生理记录。该技术将首次直接测量机电转换过程中可能发生的组织硬度的动态变化，并产生适应。与公共卫生的相关性。遗传性和后天性疾病（包括艾滋病和糖尿病）以及化疗药物都会降低触觉。据估计，此类周围感觉神经病的并发症每年造成的医疗费用超过 40 亿美元。尽管如此，人们对触觉及其在疾病中的退化仍知之甚少。在这项工作过程中开发的研究工具有可能显着增进我们的理解。