Single-nuclei transcriptome of enteric neurons in α-synuclein overexpressing mice

Overexpression of the neuronal protein α-synuclein (αSyn) is a risk factor for Parkinson’s disease (PD), and αSyn aggregation promotes neurodegeneration and movement disorders. αSyn overexpressing mice (Thy1-ASO) reproduce many PD-associated phenotypes such as motor deficits and constipation, which impacts a majority of patients. We sought to gain insight into enteric nervous system function in Thy1-ASO mice by isolating neuronal nuclei from mouse intestines, which makes up less than 1% of cells, followed by transcriptomic analysis.

To overcome the challenge of profiling gene expression in rare cell populations within complex tissues, we engineered an adeno-associated virus (AAV) that expresses a fluorescent protein in the nuclear membrane under the control of a constitutive neuronal promoter. We generated a high-quality single-nuclei gene expression dataset that revealed transcriptional heterogeneity reflecting populations of 4 putative excitatory motor neuron subtypes, 2 putative inhibitory motor neuron subtypes, 3 putative interneuron subtypes, and a putative sensory neuron gene cluster.