Exploring subcellular mitochondrial heterogeneity in neurons: a multimodal approach

线粒体在许多基本的生物过程中发挥作用，包括细胞能量的产生，这些作用已知在人体具有特定功能需求的不同区域有所不同。尽管如此，不同细胞类型的线粒体如何调节它们的遗传和转录过程以促进细胞环境的广泛需求还不完全清楚。此外，线粒体功能的异质性在单细胞水平上可能更加重要，在单细胞水平上，单个细胞的不同部分的能量需求随着时间的推移而波动。神经元具有高度复杂和极化的结构，在细胞体、轴突和突触中需要非常不同的能量，需要能量才能释放神经递质与周围的神经元交流，这肯定是神经元的情况。该项目将在单细胞和亚细胞水平上解开线粒体的异质性，以更好地了解人脑中的线粒体功能障碍。为此，我们将重点研究单个神经元线粒体的转录输出，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，并将通过在微流体设备中培养人IPSC来源的神经元来促进这一过程，从而使不同的细胞隔室(轴突、树突和胞体)能够物理上分离，并分别分析线粒体。此外，还可以使用纳米除草器技术提取单个线粒体并进行单独分析。这也将得到对IPSC衍生神经元的遗传操作的支持，以测试线粒体的功能输出。总而言之，我们的目标是识别在亚细胞水平上影响线粒体功能的特殊转录程序，并了解这些过程与大脑高级功能的相关性。

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.