The role of adaptor proteins in endosomal sorting during ultrafast endocytosis

接头蛋白在超快内吞过程中内体分选中的作用

基本信息

批准号：
10232091
负责人：
Kevin Jonathan Kruse
金额：
$ 6.64万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10232091
关键词：
Actin-Binding Protein Actins Adaptor Signaling Protein Alleles Alzheimer&apos s Disease Animals Ataxia Auxins Binding Biochemical Genetics Biological Biological Process CRISPR/Cas technology Caenorhabditis elegans Cell membrane Cells Clathrin Clathrin Adaptor Protein Complexes Color Complex Cytoskeleton Cytosol Defect Diffuse Electron Microscopy Endocytic Vesicle Endocytosis Endosomes Exhibits Exposure to FASTK Gene Failure Fellowship Fluorescence Freezing Genes Genetic Goals Hippocampus (Brain)Hot Spot Huntington Disease Location Lysosomes Mediating Membrane Methods Microscopy Morphology Mus Mutation Natural regeneration Nematoda Neurodegenerative Disorders Neuromuscular Junction Neurons Neurotransmitter Receptor Parkinson Disease Pathway interactions Play Process Proteins Recovery Recycling Resolution Role Sampling Scaffolding Protein Site Sorting - Cell Movement Specificity Synapses Synaptic Vesicles Syndrome Time Tissues Transcription Factor AP-1 Vesicle enhancer-binding protein AP-2 genetic approach intersectin 1 macromolecule mutant nano neurotransmission overexpression pleiotropism receptor synaptic function trafficking

项目摘要

Summary Endocytosis is a process by which macromolecules, receptors, transporters, and channels are recycled via internalization of the plasma membrane. Endocytosis at synapses, called synaptic vesicle endocytosis, supports the rapid recovery of vesicles during neurotransmission. Clathrin-mediated endocytosis occurs ~ 30 s after stimulation, however using “flash-and-freeze” electron microscopy we have demonstrated endocytosis at 30 – 300 ms after stimulation in both mouse hippocampal neurons as well as C. elegans neuromuscular junctions. Further, the process is clathrin independent. Ultrafast endocytosis generates a large endosome, which much be sorted into new synaptic vesicles or targeted for degradation in the lysosomes. Sorting of the endosome requires clathrin, but it is not clear which proteins act on the endosome to target these vesicles. The clathrin binding adaptor proteins AP1, AP2, and AP3 have all been implicated in synaptic function and targeting of membrane bound compartments. AP2 is thought to regenerate synaptic vesicles, while AP1 and AP3 are thought to target vesicles to the plasma membrane or the lysosome, respectively. The goal of this fellowship is to determine the role of these adaptor proteins in endosomal sorting and regeneration of synaptic vesicles during ultrafast endocytosis.

概括内吞作用是大分子、受体、转运蛋白和通道通过质膜的内吞作用进行回收。突触，称为突触小泡内吞作用，支持小泡的快速恢复在神经传递过程中，网格蛋白介导的内吞作用发生在刺激后约 30 秒，然而，使用“闪冻”电子显微镜我们已经证明刺激后 30 – 300 ms 小鼠海马神经元也发生内吞作用此外，该过程与网格蛋白无关。超快的内吞作用产生一个大的内体，它被分类成新的突触小泡或内体分选的靶向降解。需要网格蛋白，但尚不清楚哪些蛋白质作用于内体以靶向这些蛋白网格蛋白结合接头蛋白 AP1、AP2 和 AP3 均已被分离。 AP2 涉及突触功能和膜结合区室的靶向。据认为可以再生突触小泡，而 AP1 和 AP3 则被认为以小泡为目标分别针对质膜或溶酶体。确定这些衔接蛋白在内体分选和再生中的作用超快内吞作用期间的突触小泡。