Exploring subcellular mitochondrial heterogeneity in neurons: a multimodal approach

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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.