single-cell RNA sequencing on Drosophila melanogaster brain in an ALS/FTD model
The mutation in the C9orf72 gene with a hexanucleotide repeat has been reported multiple times to be the most common genetic cause of FTD and ALS (Frontotemporal Dementia and Amyotrophic Lateral Sclerosis), both of which are devastating neurodegenerative diseases having no cures currently. Our lab previously created fruit fly models expressing 36(C4G2) repeats, these are highly toxic to adult neurons of fruit flies. This is one of the most commonly used fly models of disease. Like many neurodegenerative diseases, FTD and ALS display selective neuronal vulnerability: only some neuronal populations succumb to disease, even though the toxic species are ubiquitously expressed. Our lab proposes to identify which neuronal populations are selectively depleted in response to the expression of the repeats and analyse which pathways are activated in vulnerable and resistant neuronal populations using our fly model of disease. This is done by scRNA sequencing across multiple time points, tracking disease development.  The workflow was first having the flies ready and their brains being dissected. The brains were then dissociated by collagenase and dispase, and the cell suspensions were passed through a 10um cell strainer. The single-cell suspensions were checked for viability and the single-cell libraries were prepared with 10X Chromium 3' platform.