Exploring subcellular mitochondrial heterogeneity in neurons: a multimodal approach

探索神经元亚细胞线粒体异质性：多模式方法

• 批准号: 2725880
• 金额: --
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2725880
• 关键词: Exploring; subcellular; mitochondrial; heterogeneity; neurons

Mitochondria play a role in many fundamental biological processes, including thegeneration of cellular energy, and these roles are known to vary across differentregions of the human body that have specific functional requirements. Despite this, itis not fully known how mitochondria in different cell types modulate their genetic andtranscriptional processes to facilitate a wide range of demands from the cellularenvironment. Furthermore, heterogeneity in mitochondrial function may be even moreimportant at the level of a single cell, where different compartments of an individualcell fluctuate in their energy demands over time. This is certainly the case withneurons, which have a highly complicated and polarised structure, and have verydifferent energy requirements in the cell body, along axons and at synapses whereenergy is required to release neurotransmitters to communicate with surroundingneurons. This project will disentangle mitochondrial heterogeneity at the single celland subcellular level to better understand mitochondrial dysfunction in the humanbrain. To do this, we will focus on the transcriptional output of mitochondria in singleneurons, as well as for single mitochondria across different regions of a cell. This willinvolve the computational integration of high-throughput RNA sequencing and geneticdata to characterise nuclear-mitochondrial interactions at ever higher resolutions, andwill be facilitated by culturing human iPSC-derived neurons in microfluidic devices,such that different cellular compartments (axons, dendrites and soma) can bephysically separated, and mitochondria within seperately analysed. Additionally singlemitochondria can be extracted using nanotweezer technology and individuallyanalysed. This will also be supported by genetic manipulation of iPSC-derived neuronsto test mitochondrial functional outputs. In all, we aim to identify specialisedtranscriptional programmes that influence mitochondrial function on the sub-cellularscale, and to understand the relevance of these processes to the higher-order functionof the brain.

线粒体在许多基本生物过程中发挥着作用，包括细胞能量的产生，并且已知这些作用在具有特定功能要求的人体不同区域中有所不同。尽管如此，我们还不完全了解不同细胞类型中的线粒体如何调节其遗传和转录过程，以满足细胞环境的广泛需求。此外，线粒体功能的异质性在单个细胞水平上可能更为重要，其中单个细胞的不同区室的能量需求随着时间的推移而波动。神经元的情况确实如此，神经元具有高度复杂和极化的结构，并且在细胞体、轴突和突触处具有非常不同的能量需求，其中需要能量来释放神经递质以与周围的神经元进行通信。该项目将在单细胞和亚细胞水平上解开线粒体异质性，以更好地了解人脑中的线粒体功能障碍。为此，我们将重点关注单个神经元中线粒体的转录输出，以及细胞不同区域的单个线粒体的转录输出。这将涉及高通量 RNA 测序和遗传数据的计算集成，以更高的分辨率表征核-线粒体相互作用，并将通过在微流体装置中培养人类 iPSC 衍生的神经元来促进，这样不同的细胞区室（轴突、树突和体细胞）可以物理分离，线粒体可以单独分离。 分析了。此外，可以使用纳米镊子技术提取单个线粒体并进行单独分析。这也将得到对 iPSC 衍生神经元进行基因操作以测试线粒体功能输出的支持。总之，我们的目标是确定在亚细胞尺度上影响线粒体功能的专门转录程序，并了解这些过程与大脑高级功能的相关性。