DUAL ENZYME-BASED MICROBIOSENSOR TO MEASURE ATP RELEASE FROM THE CAROTID BODY

基于双酶的微生物传感器测量颈动脉体的 ATP 释放

• 批准号: 7261771
• 负责人: ESTELLE B. GAUDA
• 金额: $ 23.94万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7261771
• 关键词: Adenosine; Adenosine Triphosphate; Affect; Aging; Binding; Biomedical Engineering; Biosensor; Breathing; Caliber; Carbon Dioxide; Carotid Body; Cells; Chemoreceptors; Collaborations; Condition; Detection; Development; Disease; Ensure; Environmental air flow; Enzymes; Gated Ion Channel; Gene Proteins; Glucose; Grant; Health; Hypoxia; In Vitro; Institutes; Ion Channel Gating; Laboratories; Lead; Life; Measurement; Measures; Mediating; Modification; Molecular; NIH Program Announcements; Nerve; Neuraxis; Neurons; Neurotransmitters; Newborn Infant; Oxygen; Oxygen measurement, partial pressure, arterial; P2X-receptor; Pathology; Peripheral; Peripheral Nervous System; Phenotype; Physiological; Physiology; Preparation; Process; Range; Rattus; Research; Research Project Grants; Resolution; Response to stimulus physiology; Senior Scientist; Sensory; Signal Transduction; Solutions; Synaptic Transmission; System; Technology; Testing; Time; Tissues; USA Georgia; Week; Western Asia Georgia; base; mature animal; millisecond; miniaturize; nanobiosensor; neurochemistry; novel; performance tests; postnatal; protein expression; prototype; receptor function; research study; response; sensor; small molecule; tool; tripolyphosphate

DESCRIPTION (provided by applicant): Neurochemicals modify the electrical activity of cells or neurons, induce gene and protein expression, which lead to short and long-term modifications in the phenotype of the neuron or cell. Thus, defining the neurochemical profile of specific cells and neurons is essential to understanding physiological and pathological functions that underlie normal development and aging in health and disease. Recent advances in micro and nanobiosensing technologies has provided an avenue to measure real-time release of neurochemical released from neurons and cells. ATP is released from neurons in the central and peripheral nervous system, and mediates fast-synaptic transmission via binding to P2X ion-gated channels. The peripheral arterial chemoreceptors in the carotid body are critical in regulating ventilation and circulatory responses to changes in oxygen tension, and they provide essential sensory input during early development to stabilize and maintain breathing throughout life. Maturation of hypoxic chemosensitivity occurs within in the first several weeks of mammalian postnatal development. We hypothesize that increase release of ATP from oxygen sensing cells in the peripheral arterial chemoreceptors may operative in mediating the development of hypoxic chemosensitivity. In response to the program announcement (PA, 03-058) for exploratory/developmental bioengineering research grants (EBRG), we aim to 1) further develop a novel dual enzyme-based amperometric microbiosensor (tip diameter 5-10 mcirons) that can measure ATP release across a wide range of physiological conditions in solution, and 2) measure ATP release from the superfused rat carotid body thereby establishing the stimulus-response profile of ATP release from the carotid body during postnatal maturation. These studies will be done in collaboration with colleagues at Georgia Institutute of Technology, who have already developed the prototype of the microbiosensor to be used in experiments outlined in this proposal. Since synaptic transmission is the core elementary process in the function of the peripheral and central nervous system, and ATP is an important molecule involved in this process, we believe that successful fabrication of this microbiosenor will allow for general application to multiple experimental paradigms that interrogates cellular and molecular mechanisms that meditate synaptic transmission during development and aging. Adenosine triphosphate (ATP) is an essential molecule that allows nerves to communicate with each other. With the correct tools, (microbiosensors) we can measure this small molecule as it is released from tissues and neurons. This grant proposes to develop a microbiosensor to measure the release of ATP from the carotid body to understand how the newborn develops an adequate breathing response to changes in oxygen concentration.

描述（由申请人提供）：神经化学物质修饰细胞或神经元的电活动，诱导基因和蛋白质表达，从而导致神经元或细胞表型的短期和长期修饰。 因此，定义特定细胞和神经元的神经化学谱对于理解健康和疾病正常发育和衰老的生理和病理功能至关重要。 微观和纳米传感技术的最新进展为测量神经元和细胞释放的神经化学的实时释放提供了途径。 ATP从中央和周围神经系统中的神经元释放，并通过与P2X离子门控通道结合介导快速突触传播。 颈动脉体中的外周动脉化学感受器对于调节通气和对氧气张力变化的循环反应至关重要，并且它们在早期发育过程中提供了必要的感​​觉输入，以稳定和维持一生的呼吸。 低氧化学敏感性的成熟发生在哺乳动物产后发育的前几周。 我们假设增加了外周动脉化学感受器中ATP从氧气传感细胞中释放的释放，可以在介导低氧化学敏感性的发展方面起作用。 为了响应计划公告（PA，03-058），用于探索性/发育生物工程研究补助金（EBRG），我们的目标是1）进一步开发一种基于双酶的新型双酶微动物（TIP直径5-10 mcirons）（可以在跨型物质条件下均可避免ATP的范围，并从解决方案范围内释放ATP，并释放了2次，并释放了2次，并释放了2次，从而从解决方案中释放了ATP，并且产后成熟过程中ATP从颈动脉体中释放的刺激反应曲线。 这些研究将与佐治亚州技术研究所的同事合作进行，他们已经开发了微生物传感器的原型，该原型旨在用于本提案中概述的实验中。 由于突触传播是周围神经系统和中枢神经系统功能的核心基本过程，并且ATP是参与此过程的重要分子，因此我们认为，该微生物的成功制造将允许在多种实验范式中进行一般应用，从而质疑细胞和分子机制，从而在发育过程中冥想突触传播。 三磷酸腺苷（ATP）是一个必不可少的分子，可以使神经相互通信。 使用正确的工具（微生物传感器），我们可以在从组织和神经元中释放出该小分子。 该赠款建议开发一种微生物传感器来测量ATP从颈动脉体的释放，以了解新生儿如何对氧气浓度变化产生适当的呼吸反应。