MECHANISM FUSION TRANSMITTER RELEASE IN NEUROSECRETION

描述（改编自申请人摘要）：长期原则 该项目的目标是提供一个详细的生物物理和分子 对内分泌细胞胞吐囊泡融合和递质释放的认识 细胞和神经末梢。在电刺激神经末梢时， 内分泌细胞释放多种神经递质和神经肽， 胞吐机制这种神经分泌的过程是非常显著的 重要性在广泛的生理过程。详细 因此，对胞吐事件的理解具有实质性的意义， 特别是对于那些神经分泌受损的疾病。在胞吐作用中 囊泡停靠在质膜上并与质膜融合 允许囊泡内容物通过融合孔释放。是 建议研究个别融合事件和个别对接事件， 生物物理技术聚变和释放的同时测量将 通过“贴片安培法”进行，该方法允许同时 记录微小的电容变化和安培检测释放的 传递分子。“细胞附着”贴片的实验提供了 关于胞质条件和组分的要求的信息。到 研究被认为在体外融合之前的对接过程， 申请人将开发一种使用光阱的新方法。他们将 表征决定对接和发射器的物理化学机制 release.这些方法将与生物化学技术相补充， 将用于表征特定融合孔形成的作用（ 吻和运行机制）或可替代地通过完全融合， 通过包被的小凹进行常规的内吞作用。长期目标是获得 了解直接参与融合孔的分子组分 阵证明重组系统中的融合孔 类似于胞吐过程中的融合孔形成， 其意义类似于证明特异性膜蛋白 在脂质膜中重构的类似于特定离子的性质 渠道

DESCRIPTION(Adapted from applicant's abstract): The principle 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 processes. A detailed understanding of the exocytotic event is thus of substantial interest, in particular for those diseases where neurosecretion is impaired. In exocytosis the vesicles dock at the plasma membrane and fuse with the plasma membrane allowing for release of the vesicle contents through the fusion pore. It is proposed to study individual fusion events and individual docking events using biophysical techniques. Simultaneous measurements of fusion and release will be performed by "patch amperometry", a method which allows simultaneous recording of small capacitance changes and amperometric detection of released transmitter molecules. Experiments in "cell-attached" patched provide information on the requirements of cytosolic conditions and components. To investigate the docking process which is thought to precede fusion in vitro, the applicant will develop a novel approach using an optical trap. They will characterize the physicochemical mechanisms determining docking and transmitter release. These methods will be complemented with biochemical techniques which will be used to characterize the role of specific fusion pore formation (the kiss-and-run mechanism) or alternatively by full fusion followed by conventional endocytosis via coated pits. The long-term aim is to obtain an understanding of the molecular components directly involved in fusion pore formation. Demonstration that the fusion pore in a reconstituted system resembles the fusion poreforming during exocytosis will eventually be of a significance analogous to that demonstrating that specific membrane proteins reconstituted in lipid membranes resemble the properties of specific ion channels.