Investigating the transcriptional and functional stress responses in different neuron subtypes after knockdown of mitochondrial complex I subunit ND75

We hypothesise that mitochondrial complex I inhibition (via knock down of subunit ND-75) will lead to distinct transcriptional and functional stress responses in different neuronal subtypes. To test this hypothesis in the next upcoming months I plan to:

1. Analyze the behavioural effects of ND-75 KD in a single neuronal subtype, dopaminergic and serotonergic neurons.
Climbing and seizure assays
CAFÉ feeding assay
ANY-Maze open field assay
Hot plate assay
Analysis of ATF4 expression- adult brain dissections, staining and confocal microscopy

2. Analyze the transcriptional response caused by ND-75 KD in single neurons.
-We will perform single nuclear RNA-seq (snRNA-seq) from flies with pan-neuronal ND-75 KD and ND-75 KD together with ATF4 KD.

3. Test whether targeting mitochondrial stress signalling modifies dopaminergic and serotonergic neuron function.