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）：神经化学物质改变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改变。因此，定义特定细胞和神经元的神经化学特征对于理解健康和疾病中正常发育和衰老的生理和病理功能至关重要。微纳米生物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物质的实时释放提供了途径。ATP从中枢和外周神经系统的神经元释放，并通过结合P2X离子门控通道介导快速突触传递。颈动脉体中的外周动脉化学感受器在调节通气和对氧张力变化的循环反应中至关重要，它们在早期发育过程中提供必要的感觉输入，以稳定和维持生命中的呼吸。缺氧化学敏感性的成熟发生在哺乳动物出生后发育的最初几周内。我们假设外周动脉化学感受器中的氧感应细胞释放ATP的增加可能在缺氧化学敏感性的发展中起作用。为了响应探索性/发育性生物工程研究资助（EBRG）的项目公告（PA, 03-058），我们的目标是：1)进一步开发一种新型的基于双酶的安培微生物传感器（尖端直径5-10微米），可以测量溶液中多种生理条件下ATP的释放；2)测量过量的大鼠颈动脉体释放ATP，从而建立出生后成熟过程中颈动脉体释放ATP的刺激-反应谱。这些研究将与乔治亚理工学院的同事合作完成，他们已经开发出微生物传感器的原型，用于本提案中概述的实验。由于突触传递是外周和中枢神经系统功能的核心基本过程，而ATP是参与这一过程的重要分子，我们相信这种微生物传感器的成功制造将允许广泛应用于多种实验范式，以询问在发育和衰老过程中冥想突触传递的细胞和分子机制。三磷酸腺苷（ATP）是神经相互沟通的重要分子。有了正确的工具（微生物传感器），我们可以测量这种小分子从组织和神经元中释放出来的情况。这项拨款建议开发一种微生物传感器来测量颈动脉体ATP的释放，以了解新生儿如何对氧浓度的变化产生适当的呼吸反应。