MEASURING FASTER PHYSIOLOGICAL EVENTS WITH ELECTROCHEMICAL PROBES

使用电化学探针更快地测量生理事件

• 批准号: 7598479
• 负责人: MARK A MESSERLI
• 金额: $ 7.03万
• 依托单位: MARINE BIOLOGICAL LABORATORY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-12-01 至 2007-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7598479
• 关键词: Blur; Calcium-Activated Potassium Channel; Cell physiology; Cells; Chemicals; Chinese Hamster Ovary Cell; Computer Retrieval of Information on Scientific Projects Database; Condition; Data Analyses; Data Collection; Detection; Diffuse; Electrodes; Environment; Event; Funding; Gases; Grant; Individual; Institution; Invasive; Ion Channel; Ion-Selective Electrodes; Ions; Lipid Bilayers; Measures; Metabolic; Methods; Microelectrodes; Monitor; Nature; Noise; Physiological; Range; Reaction Time; Regulation; Research; Research Personnel; Resources; Scanning; Scheme; Signal Transduction; Source; Speed; Surface; System; Techniques; Tissues; United States National Institutes of Health; Xenopus oocyte; data modeling; design; insight; response; sensor; uptake; wasting

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Cells maintain a dynamic interaction with their environment by acquiring and expelling inorganic ions, gases and organic compounds ranging from metabolic wastes to chemical messengers. Release of a compound results in a surface high concentration that produces a gradient as the compound diffuses away from the cell; uptake results in an inverse gradient. These gradients are measured using modulation of electrochemical probes that enhance the signal to noise ratio but to date have been restricted to relatively steady state applications. Studies conducted at the BRC reviewed the expected response time for electrochemical sensors and noted that several of the potentiometric design can achieve 90% response in less than 20msec. This speed brings us to within the scope of measuring channel activities. We have used K+-selective microelectrodes to monitor changes in external [K+] after efflux through artificial channels in a planar lipid bilayer and after efflux through Ca2+-activated K+ channels expressed in Xenopus oocytes and Chinese Hamster Ovary cells. A combination of modeling and data analysis schemes has been used to confirm single channel detection and identify the strengths and weaknesses of the system. Combining these non-invasive sensors and analysis approaches with a scanning technique described elsewhere will provide a unique insight into cellular organization, revealing finer details of spatial and temporal regulation of cellular processes from chemical gradients surrounding cells. Self-referencing with ion-selective electrodes (ISEs) has been used noninvasively, to measure relatively steady ionic gradients near cells and tissues. However, these relatively steady gradients are the average of many discrete events including transport through channels or transporters. The data collection and averaging scheme used for measuring the steady gradients blurs the individual events, leading to the loss of useful information regarding the nature of the ionic gradient. By using fast responding electrodes with signal analysis methods we hope to characterize ion channels and transporters under normal and pathogenic conditions in order to study the diseased state with a non-invasive approach.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 细胞通过获取和驱除无机离子，气体和有机化合物的动态相互作用，从代谢废物到化学信使。化合物的释放会导致表面高浓度，该浓度会产生梯度，因为该化合物从细胞中扩散。摄取导致反向梯度。这些梯度是使用电化学探针的调节来测量的，该探针增强了信号与噪声比，但迄今已限于相对稳态的应用。 在BRC进行的研究回顾了电化学传感器的预期响应时间，并指出，几种电位测量设计可以在小于20msec的情况下实现90％的响应。这种速度使我们进入了测量渠道活动的范围。我们已经使用K+选择性微电极通过在平面脂质双层中的人造通道中进行外部[K+]的变化，并在通过Ca2+激活的K2+激活的K+通道中排出后，以异武型卵母细胞和中国仓鼠的羊仓卵巢细胞表达。 建模和数据分析方案的结合已被用来确认单个通道检测并确定系统的优势。 将这些非侵入性传感器和分析方法与其他地方描述的扫描技术相结合，将为细胞组织提供独特的见解，从而揭示了从周围化学梯度周围化学梯度的空间和时间调节细胞过程的细节。 用离子选择电极（ISE）自我引用已无创使用，以测量细胞和组织附近相对稳定的离子梯度。 但是，这些相对稳定的梯度是许多离散事件的平均值，包括通过通道或转运蛋白运输。 用于测量稳定梯度的数据收集和平均方案模糊了单个事件，从而导致有关离子梯度性质的有用信息的丢失。 通过使用信号分析方法的快速响应电极，我们希望在正常和致病条件下表征离子通道和转运蛋白，以便以非侵入性方法研究患病状态。