Force Clamp Systems for Evaluation of Mechanotransduction

用于评估机械传导的力夹系统

• 批准号: 8244400
• 负责人: Miriam B Goodman
• 金额: $ 46.68万
• 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 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8244400
• 关键词: Accounting; Acquired Immunodeficiency Syndrome; Address; Affect; Aging; American; Amputation; Animal Model; Animals; Award; Beds; Behavior; Behavioral; Biological Assay; Biological Process; Body measure procedure; Caenorhabditis elegans; Caring; Cells; Cytoskeleton; Defect; Devices; Diabetes Mellitus; Diagnostic; Disease; Dissection; Engineering; Evaluation; Exhibits; Extracellular Matrix; Eyebrow structure; Functional disorder; Generations; Genetic; Genetic Models; Goals; HIV; Hair; Healed; Health Care Costs; Hydrostatic Pressure; Image; In Vitro; Indium; Individual; Inherited; Injury; Intervention; Ion Channel; Ion Channel Protein; Kinetics; Knowledge; Lead; Learning; Life; Limb structure; Location; Lower Extremity; Mammals; Maps; Measurement; Measures; Mechanical Stimulation; Mechanics; Mechanoreceptors; Mediating; Medical; Methods; Metric; Modeling; Molecular; Muscle; Muscle Contraction; Muscle Tonus; Mutation; Nematoda; Nerve Endings; Nervous system structure; Neurons; Optics; Pacinian Corpuscles; Pain; Pathology; Pathway interactions; Peripheral Nervous System Diseases; Phenotype; Positioning Attribute; Preparation; Process; Property; Proprioception; Proteins; Protocols documentation; Public Health; Quality of life; Research; Risk Factors; Role; Running; Sensory; Sensory Nerve Endings; Skin; Stimulus; Structure; Subcellular structure; Subcutaneous Tissue; System; Testing; Time; Touch sensation; Work; analytical method; animal data; base; cantilever; chemotherapy; design; diabetic; healing; improved; in vitro Model; in vivo; microsystems; model development; mutant; optogenetics; patch clamp; pressure; protein structure; public health relevance; ratiometric; receptor; research study; response; sensor; sensory neuropathy; social communication; tool; transmission process

DESCRIPTION (provided by applicant): The long-term goal of this research is to discover the force transmission and force transduction pathways responsible for touch and proprioception. These senses are essential for social communication and every aspect of daily life from sitting, to standing, to running; however, their function is disrupted in both inherited and acquired diseases, including HIV-AIDS and diabetes. Even partial loss of sensory function as in diabetic peripheral neuropathy (DPN) has devastating consequences; DPN affects an estimated 15 million Americans and is the dominant risk factor in lower limb amputations. Thus, the loss of touch and proprioception is common, associated not only with discomfort and pain, but also with a decrease in the quality of life. Despite this, diagnostic tools and treatments for the dysfunction of touch and proprioception remain poorly developed, principally because little is known about how these senses work. This knowledge gap reflects a lack of adequate devices for delivering controlled mechanical stimuli and of animal models amenable to analysis of the mechanobiology of touch sensation. The objective of the proposed research is to bridge this gap by developing new devices for controlled force delivery, improved animal models for dissecting force transmission and transduction pathways, and new analytical methods for fundamental study of the relevant mechanics of this basic life process. The proposed research uses the simple roundworm, Caenorhabditis elegans, because more is understood about its sense of touch than that of any other animal. Research using C. elegans has successfully revealed mechanistic aspects of several fundamental and conserved biological processes, including touch sensation. It was in C. elegans, for instance, that the first ion channel proteins required for touch sensation were identified ~20 years ago. Because analogous proteins are expressed in mammalian touch receptor neurons, they may also contribute to touch sensation. At present, C. elegans is the only animal in which we know which proteins form the mechano-electrical transduction channels responsible for detecting force in touch receptor neurons. This knowledge enables a level of analysis that is not currently available in mammalian models. The central hypothesis we are testing is that both force-sensitivity and response dynamics are determined by the interplay of skin mechanics, neuron position, and intracellular, cytoskeletal structures. To test this hypothesis, we will develop new metrics for quantitative assessment of touch sensitivity; new microfabricated tools suitable for delivering pN-5N forces, new in vitro models of touch receptor neurons, and build new models of force transmission and force transduction. The specific aims are: 1) Test the hypothesis that skin mechanics, neuron position, and the neuronal cytoskeleton regulate touch sensitivity in vivo; 2) Assess the impact of body wall muscle tone and internal hydrostatic pressure on touch sensation in vivo; 3) Identify mechanisms of mechano- electrical transduction channel activation and adaptation. PUBLIC HEALTH RELEVANCE: Normal touch sensation and proprioception are essential for daily life and require the activation of specialized mechanoreceptor neurons. When the function of such neurons is disrupted by aging, disease (HIV-AIDS, diabetes), and medical interventions (chemotherapy), small injuries often lead to wounds that fail to heal and are treated only by limb amputation. Such pathologies afflict millions of Americans and account for tens of billions of dollars in health-care costs annually. By revealing the mechanisms by which force is transmitted from the skin to mechanoreceptor neurons and developing microfabricated tools for research, these studies may provide new strategies for 1) interventions that could restore sensitivity in individuals afflicted by sensory neuropathy and for 2) improved diagnostic tools that may aid in the application of interventions that minimize the loss of sensory function.

描述（由申请人提供）：本研究的长期目标是发现负责触摸和本体感受的力传递和力转导途径。这些感觉对于社交和日常生活的各个方面都是必不可少的，从坐到站，再到跑步;然而，它们的功能在遗传性和后天性疾病中都被破坏，包括艾滋病和糖尿病。即使是部分感觉功能丧失，如糖尿病周围神经病变（DPN），也会产生毁灭性的后果;据估计，DPN影响了1500万美国人，是下肢截肢的主要危险因素。因此，触觉和本体感觉的丧失是常见的，不仅与不适和疼痛有关，而且与生活质量的下降有关。尽管如此，触觉和本体感觉功能障碍的诊断工具和治疗方法仍然很不发达，主要是因为对这些感觉如何工作知之甚少。这种知识差距反映了缺乏足够的设备提供控制的机械刺激和动物模型服从的机械生物学的触觉分析。拟议的研究的目的是通过开发新的设备，控制力传递，解剖力传递和转导途径的动物模型，和新的分析方法，这个基本的生命过程的相关力学的基础研究，以弥合这一差距。这项拟议中的研究使用了简单的蛔虫，秀丽隐杆线虫，因为它的触觉比其他任何动物都要多。使用C.秀丽隐杆线虫已经成功地揭示了几种基本的和保守的生物过程的机制方面，包括触觉。是C调的。例如，线虫，第一个触觉所需的离子通道蛋白在大约20年前就被发现了。因为类似的蛋白质在哺乳动物触觉感受器神经元中表达，它们也可能有助于触觉。目前，C.我们知道，在线虫动物中，哪种蛋白质形成了负责检测触觉感受器神经元中的力的机械-电传导通道。这一知识使得目前在哺乳动物模型中不可用的分析水平成为可能。我们正在测试的中心假设是，力敏感性和响应动力学都是由皮肤力学、神经元位置和细胞内细胞骨架结构的相互作用决定的。为了验证这一假设，我们将开发新的指标，用于定量评估触摸敏感性;新的微加工工具，适用于提供pN-5 N力，新的触摸感受器神经元体外模型，并建立新的力传递和力转导模型。具体目标是：1）测试皮肤力学、神经元位置和神经元细胞骨架在体内调节触觉敏感性的假设; 2）评估体壁肌张力和内部流体静压对体内触觉的影响; 3）识别机械-电转导通道激活和适应的机制。 公共卫生关系：正常的触觉和本体感觉对于日常生活是必不可少的，并且需要激活专门的机械感受器神经元。当这些神经元的功能被衰老、疾病（艾滋病、糖尿病）和医疗干预（化疗）破坏时，小的损伤往往会导致伤口无法愈合，只能通过截肢来治疗。这些疾病折磨着数百万美国人，每年造成数百亿美元的医疗保健费用。通过揭示力从皮肤传递到机械感受器神经元的机制，并开发用于研究的微制造工具，这些研究可能为以下方面提供新的策略：1）可以恢复受感觉神经病折磨的个体的敏感性的干预措施，以及2）改进的诊断工具，可以帮助应用干预措施，最大限度地减少感觉功能的丧失。