Mechanisms of Fusion & Transmitter release

融合机制

• 批准号: 7255486
• 负责人: Manfred LINDAU
• 金额: $ 27.72万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-20 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7255486
• 关键词: Address; Affect; Amino Acids; Behavior; Binding; Biochemical; Biological Models; Brain; C-terminal; Catecholamines; Cell membrane; Cells; Charge; Chromaffin Cells; Cisplatin/Doxorubicin/Melphalan/Teniposide; Complex; Cytoplasmic Granules; Cytoplasmic Vesicles; Detection; Dimerization; Disease; Dissociation; Docking; Electric Capacitance; Electric Stimulation; Emotional; Endocrine; Endocytosis; Endopeptidases; Equus caballus; Event; Exocytosis; Fluorescence Resonance Energy Transfer; Hormones; Human body; Image; In Vitro; Individual; Length; Location; Measurement; Mechanics; Mediating; Mediator of activation protein; Membrane; Membrane Fusion; Membrane Proteins; Methods; Microscopy; Molecular; Mutation; Nerve; Neuromodulator; Neurons; Neuropeptides; Neurosecretion; Neurotransmitters; Object Attachment; Peptide Hydrolases; Peripheral; Permeability; Phosphorylation; Physiological; Physiological Processes; Process; Property; Protein Overexpression; Proteins; Range; Rate; Regulation; Reporting; Research Personnel; Role; Running; SNAP receptor; Secretory Vesicles; Son; Synaptic Transmission; Techniques; Testing; Tetradecanoylphorbol Acetate; Transmembrane Domain; Vesicle; Width; Work; base; carbon fiber; cell type; eosinophil; granulocyte; human VAPA protein; innovation; interest; laser tweezer; mutant; programs; research study; response; size; syntaxin; vesicle-associated membrane protein

DESCRIPTION (provided by applicant): The principal long-term objective of this project is to provide a detailed biophysical and molecular understanding of exocytotic vesicle fusion and transmitter release in endocrine cells and nerve terminals. Upon electrical stimulation nerve terminals and endocrine cells release a variety of neurotransmitters and neuropeptides by an exocytotic mechanism. This process of neurosecretion is of outstanding importance in a broad range of physiological functions in the human body. It allows for synaptic transmission as well as release of hormones and neuromodulators and is thus an essential event mediating brain function, emotional response, behavior and various other physiological processes. A detailed understanding of the exocytotic event is of substantial interest, in particular for those diseases where neurosecretion is impaired. In exocytosis the vesicles are tethered at the plasma membrane and fuse with the plasma membrane allowing for release of the vesicle contents through the fusion pore. This project addresses 3 specific questions: 1) How is the mode of exocytosis (kiss-and run or full fusion) regulated? 2) What is the mechanism of fusion pore formation? 3) How are vesicles tethered to their target membrane before fusion is stimulated? We will investigate individual fusion events and individual tethering events using biophysical techniques. Fusion pore properties such as initial and mean conductance, conductance fluctuations, expansion rate, lifetime and permeability for transmitter molecules will be studied in chromaffin cells by patch capacitance measurements and analyzed statistically. Simultaneous measurements of fusion and release will be performed by "patch amperometry", a method that allows simultaneous recording of small capacitance changes and amperometric detection of released transmitter molecules. Biochemical manipulations and over-expression of wild type and mutant proteins implicated in fusion and release are used to investigate the role of specific proteins in determining the properties of the fusion pore. The biophysical properties of tethers are studied employing force measurements with optical tweezers using eosinophil granules as a model system.

描述（由申请人提供）： 该项目的主要长期目的是提供对内分泌细胞和神经末端中胞胞菌囊泡融合和递质释放的详细生物物理和分子理解。电刺激后，神经末端和内分泌细胞通过胞吐机制释放了多种神经递质和神经肽。在人体的广泛生理功能中，这种神经分泌的过程至关重要。它允许突触传播以及激素和神经调节剂的释放，因此是介导大脑功能，情绪反应，行为以及各种其他生理过程的重要事件。对胞吐事件的详细理解具有重大关注，特别是对于那些神经分泌受损的疾病。在胞吐作用中，囊泡在质膜上束缚，并与质膜融合，从而可以通过融合孔释放囊泡含量。该项目解决了3个具体问题：1）胞吐作用（亲吻和奔跑或完全融合）的方式如何受到调节？ 2）融合孔形成的机制是什么？ 3）在刺激融合之前，如何将囊泡束缚在其靶膜上？我们将使用生物物理技术研究个别融合事件和个人绑扎事件。融合孔特性，例如初始电导和平均电导率，电导率波动，膨胀速率，寿命和发射机分子的渗透性将通过斑块电容测量和统计学分析在铬蛋白细胞中研究。融合和释放的同时测量将通过“斑块安培法”进行，该方法允许同时记录释放的发射机分子的小电容变化和安培检测。与融合和释放有关的野生型和突变蛋白的生化操纵和过表达用于研究特定蛋白在确定融合孔的性质中的作用。使用嗜酸性粒细胞颗粒作为模型系统，研究了使用力测量的系数的生物物理特性。

项目成果

Prostaglandin E1 inhibits endocytosis in the β-cell endocytosis.

前列腺素E1 抑制β 细胞内吞作用的内吞作用。

• DOI: 10.1530/joe-15-0435
• 发表时间: 2016
• 期刊: The Journal of endocrinology
• 作者: Zhao,Ying;Fang,Qinghua;Straub,SusanneG;Lindau,Manfred;Sharp,GeoffreyWG
• 通讯作者: Sharp,GeoffreyWG

Tethering forces of secretory granules measured with optical tweezers.

用光镊测量分泌颗粒的束缚力。

• DOI: 10.1529/biophysj.108.132670
• 发表时间: 2008
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Valero,Vicente;Nevian,Thomas;Ho,Dominik;Lindau,Manfred
• 通讯作者: Lindau,Manfred

F-actin and myosin II accelerate catecholamine release from chromaffin granules.

F-肌动蛋白和肌球蛋白 II 加速儿茶酚胺从嗜铬颗粒中的释放。

• DOI: 10.1523/jneurosci.2818-08.2009
• 发表时间: 2009-01-21
• 期刊: The Journal of neuroscience : the official journal of the Society for Neuroscience
• 作者: Berberian K;Torres AJ;Fang Q;Kisler K;Lindau M
• 通讯作者: Lindau M

Synaptobrevin-2 C-Terminal Flexible Region Regulates the Discharge of Catecholamine Molecules.

Synaptobrevin-2 C 末端柔性区域调节儿茶酚胺分子的放电。

• DOI: 10.1016/j.bpj.2019.01.028
• 发表时间: 2019
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Weiss,AnnitaN

Fusion gains independence.

融合获得独立性。

• DOI: 10.1085/jgp.200810050
• 发表时间: 2008
• 期刊: The Journal of general physiology
• 作者: Lindau,Manfred