权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

New optical approaches to study secretory granule structure and function

研究分泌颗粒结构和功能的新光学方法

基本信息

批准号：
8281418
负责人：
Daniel Axelrod
金额：
$ 11.66万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8281418
关键词：
Applications Grants Biological Cardiovascular system Catecholamines Cell membrane Cells Cellular biology Characteristics Collaborations Cytoplasm Cytoplasmic Granules Cytoskeleton Cytosol Development Disease Docking Endocrine system Event Exocytosis Fluorescence Fluorescence Microscopy Gel Goals Health Hormones Image Individual Kinetics Label Laboratories Life Measurable Measurement Measures Membrane Membrane Fusion Membrane Proteins Methods Microscope Molecular Weight Motion Nervous system structure Neurons Neurotransmitters Optics Pathway interactions Physics Physiological Play Process Proteins Research Role Secretory Cell Secretory Vesicles Shapes Site Structure Techniques Testing Time Travel Vesicle Viscosity base controlled release direct application experience insight luminal membrane novel physical state research study response

项目摘要

DESCRIPTION (provided by applicant): Our research focuses on events in the exocytotic pathway that occur in the highly specialized domain of the plasma membrane-cytoplasm interface. This region is superbly imaged by total internal reflection fluorescence microscopy (TIRFM), a core technique that we use extensively in our studies. This proposal is based upon the hypothesis that the mobility characteristics of granule lumenal contents and of granule membrane proteins shape the secretory response. The proposal will provide fundamental new insights concerning secretory granule structure and function in exocytosis and will provide the first quantitative measures of the rotational and translational mobility of granule lumenal and membrane proteins of individual granules. The rotational and translational mobility of lumenal and granule membrane proteins and of individual granules will be measured by novel combinations of TIRFM, polarization and FRAP. There are several related goals in the proposal: 1) to understand the physical state of the granule lumen and reveal the role of lumenal viscosity in determining the rates of protein and catecholamine release, 2) to determine the translational mobility of granule membrane proteins and whether the mobility permits recruitment to the fusion site on the granule membrane, 3) to determine the rotational mobility of individual granules in order to better define the tethered and/or caged state of the granules before fusion, and 4) and to determine whether the increase in granule travel immediately before fusion reflects a combination of translational and rotational motion that permits the granule to 'roll' into a fusion competent interaction with the plasma membrane.

描述(由申请人提供)：我们的研究集中在胞吐途径中发生在质膜-细胞质界面高度专门化领域的事件。这一区域由全内反射荧光显微镜(TIRFM)成像，这是我们在研究中广泛使用的核心技术。这一建议是基于这样的假设，即颗粒腔内容物和颗粒膜蛋白的流动性特征塑造了分泌反应。这项提议将提供关于分泌颗粒在胞吐作用中的结构和功能的新的基本见解，并将提供单个颗粒的颗粒、腔和膜蛋白的旋转和平移流动性的第一个定量测量。管腔和颗粒膜蛋白以及单个颗粒的旋转和平移迁移率将通过TIRFM、偏振和FRAP的新组合来测量。该提案有几个相关目标：1)了解颗粒管腔的物理状态，并揭示管腔粘度在决定蛋白质和儿茶酚胺释放速率中的作用；2)确定颗粒膜蛋白的平移迁移率以及这种迁移率是否允许重新聚集到颗粒膜上的融合部位；3)确定单个颗粒的旋转迁移率，以便更好地确定融合前颗粒的拴住和/或笼养状态；以及4)确定融合前颗粒行程的增加是否反映了平移和旋转运动的组合，使颗粒能够与质膜‘滚动’进入融合有效的相互作用。