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Analytical Techniques for Exocytosis

胞吐作用的分析技术

基本信息

批准号：
7595199
负责人：
ANDREW G EWING
金额：
$ 22.2万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-05-01 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7595199
关键词：
Ablation Area Attention Behavior Blood capillaries Brain Calcium Caliber Capillary Electrophoresis Catecholamines Categories Cell membrane Cells Cellular biology Characteristics Charge Chemistry Communication Coupled Cytolysis Data Detection Development Disease Dyes Electrochemistry Electrophoresis Environment Estrogens Event Exocytosis Fluorescence Fluorescence Microscopy Goals Homeostasis Hormones Image Journals Laboratories Lasers Lead Lipids Measurement Measures Membrane Membrane Protein Traffic Methodology Methods Modeling Molecular Nanotubes Neuromodulator Neurons Neurosciences Neurotransmitters Organelles PC12 Cells Parkinson Disease Pharmacological Treatment Pheochromocytoma Play Process Progress Reports Publishing Regulation Reporting Research Role Running Source Structure Synaptic plasticity System Techniques Technology Testing Transmembrane Transport Vesicle Work analog analytical method base capillary falls fluorescence imaging fluorophore interest nanometer neurotransmission neurotransmitter release organelle movement research study

项目摘要

The goal of the proposed research is to develop and apply analytical methods in extremely small environments to probe specific mechanisms that regulate and modulate exocytosis. A key aspect of this work will be to use amperometric measurements simultaneously with fluorescence imaging to test the hypothesis that control of calcium homeostasis and exocytosis are part of the mechanism by which estrogen is neuroprotective, to use small-volume separations to determine the extent to which the vesicle contents are released during an exocytosis event, and to use amperometric experiments and confocal fluorescence imaging to examine a new and potentially controversial hypothesis that lipid nanotubes play a role in regulating vesicle state and release. This work will be done with pheochromocytoma (PC12) cells in culture. Thus, the experiments proposed here are targeted at understanding exocytosis at the molecular level and understanding how cells are "wired" in a way that might be more general in cell biology. The specific aims of the proposal are:1) to use amperometry and calcium imaging to examine the neuroprotective effects of estrogen by measuring catecholamine release and calcium entry; 2) to develop electrophoresis with electrochemical detection in small capillaries to determine the level of neuromessenger in vesicles and by comparison to amperometric measurements at cells to determine the fraction released during exocytosis; 3) to identify lipid nanotube structures in cells by fluorescence and to investigate the mechanism of membrane trafficking to and from vesicles; and 4) to use electrochemistry and fluorescence to examine and develop models of transmitter release via the fusion pore prior to full exocytosis and an alternative hypothesis for "kiss and run"release. This proposal captures the work that we envision as necessary to truly understand the function of vesicles and exocytosis in neurotransmission and synaptic plasticity. The highly preliminary data we have obtained suggest that lipid nanotubes exist connecting vesicles to other structures, perhaps each other. If this is correct, it represents a new idea in cell biology and could be incredibly important. Overall, the use of amperometry, fluorescence and separations is proposed to investigate cellular chemistry and new ideas in cell biology related to neuronal communication.

提出的研究目标是开发和应用分析方法在极小的

项目成果

期刊论文数量（19）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Temporal resolution in electrochemical imaging on single PC12 cells using amperometry and voltammetry at microelectrode arrays.

DOI：
10.1021/ac102502g
发表时间：
2011-01-15
期刊：
ANALYTICAL CHEMISTRY
影响因子：
7.4
作者：
Zhang, Bo;Heien, Michael L. A. V.;Santillo, Michael F.;Mellander, Lisa;Ewing, Andrew G.
通讯作者：
Ewing, Andrew G.

Steady-state electrochemical determination of lipidic nanotube diameter utilizing an artificial cell model.

DOI：
10.1021/ac902282d
发表时间：
2010-02-01
期刊：
ANALYTICAL CHEMISTRY
影响因子：
7.4
作者：
Adams, Kelly L.;Engelbrektsson, Johan;Voinova, Marina;Zhang, Bo;Eves, Daniel J.;Karlsson, Roger;Heien, Michael L.;Cans, Ann-Sofie;Ewing, Andrew G.
通讯作者：
Ewing, Andrew G.

Probing the lipid chemistry of neurotoxin-induced hippocampal lesions using multimodal imaging mass spectrometry.