Mechanisms regulating mitochondrial polarity in neurons: regional distribution and specialization

项目总结： 神经退行性疾病仍然是人类最难治疗的疾病之一 理解，更不用说临床护理了。基础神经生物学的基础研究是 需要阐明疾病机制并为可能的治疗方法提供研究选择 关心。神经元是神经系统中的极化细胞，接受和传递电和 化学信息。许多细胞机制支持这种神经元功能，从 细胞骨架结构，核基因表达，到极化细胞器运输。线粒体 是在轴突和神经元的特定部位支持多种神经元功能的细胞器 并在每个隔室中保持非常不同的动态。线粒体形态 运输行为反映了线粒体在各种细胞类型中的功能和对 正常的动力学与细胞功能障碍和人类疾病有关。此外， 线粒体功能障碍是神经退行性疾病的标志。的研究。 哺乳动物海马神经元中线粒体的形态、动力学和运输将 为更好地理解线粒体网络的维持方式铺平道路 在神经元中。 这项研究将集中于确定线粒体在多大程度上被专门化 神经元亚室。使用先进的活细胞荧光技术，微流控 平台，以及下一代蛋白质组学和RNAseq分析，我们将识别分子 线粒体极性形成和维持的机制 成熟神经元中的网络。此外，文献和我们最初的见解，支持一个角色 Mitofusin 2在调节神经元线粒体网络中的作用我们正在开发仿冒品 研究Mfn2在神经元线粒体融合中的作用的技术。续 这项新技术的开发将提供概念上的证明，我们相信 一般适用于大多数膜蛋白。跨膜蛋白是一种 特殊挑战当试图彻底灭活它们时，迫切需要 仿冒品的发展。此外，来自不同亚群的单个线粒体 通过新的分类和纯化，神经元中的隔间将被物理隔离 程序。将对这些线粒体进行蛋白质组检测，以了解它们是如何 线粒体分类，以及最终线粒体网络是如何维持的。来自以下方面的见解 这些研究将被用来理解由非典型肺炎引起的神经元病理。 由胆碱激酶β(CHKB)突变引起的先天性肌营养不良。

Project Summary: Neurodegenerative diseases remain some of the most difficult human maladies to understand, much less clinically care for. Fundamental research into basic neurobiology is needed to clarify disease mechanisms and provide research options for possible therapeutic care. Neurons are polarized cells in the nervous system that receive and transmit electrical and chemical information. Many cellular mechanisms support this neuronal function from cytoskeletal structure, nuclear gene expression, to polarized organelle trafficking. Mitochondria are organelles that support a variety of neuronal functions at specialized sites in axons and dendrites and maintain very different dynamics in each compartment. Mitochondrial morphology and trafficking behavior reflect mitochondrial function in a variety cell types and disruption of normal dynamics are related to cellular dysfunction and human disease. Moreover, dysfunctional mitochondria are a hallmark of neurodegenerative diseases. The study of mitochondrial morphology, dynamics, and trafficking in mammalian hippocampal neurons will pave the way forward to a better understanding of how mitochondrial networks are maintained in neurons. This study will focus on determining to what extent mitochondria are specialized in neuronal sub-compartments. Using advanced live cell fluorescent techniques, microfluidic platforms, and next generation proteomics and RNAseq analysis, we will identify the molecular mechanisms behind the formation of mitochondrial polarity and maintenance of mitochondrial networks in mature neurons. Furthermore, literature and our initial insights, support a role for mitofusin 2 in regulating neuronal mitochondrial networks. We are developing knock-OFF technology to study the role of Mfn2 as it pertains to mitochondrial fusion in neurons. Continued development of this new technology will provide a proof of concept and we believe will be generally applicable to the majority of membrane proteins. Transmembrane proteins present a special challenge when trying to acutely inactivate them, there is an urgent need for development of knock-OFF. Furthermore, individual mitochondria from different sub- compartments in neurons will be physically isolated through novel sorting and purification procedures. These mitochondria will be assayed for their proteome to gain insight into how mitochondrial sort and ultimately how the mitochondrial network is maintained. Insights from these studies will be used to understand the neuronal pathology resulting from an atypical congenital muscular dystrophy caused by mutations in choline kinase beta (CHKB).