Transcriptional profiling of individual airway projecting vagal sensory neurons

Mammalian airways and lungs are richly innervated by bronchopulmonary sensory neurons, the vast majority of which are derived from the vagal sensory ganglia. In the present study we set out to perform high coverage single cell RNA sequencing on a population of identified murine bronchopulmonary sensory neurons collected from the vagal sensory ganglia to better define the molecular expression profiles of these cell types. Given the importance of P2X2 in differentiating nodose from jugular sensory neurons, we further aimed to investigate the relationship between transcriptional expression of identified genes and P2X2 expression. Bronchopulmonary sensory neurons in the vagal ganglia were identified by retrograde neuronal tracing with the retrograde lipophilic tracer 1,1'-Dioctadecyl-3,3,3',3'-Tetramethylindocarbocyanine Perchlorate (DiI 2%). Retrogradely labeled neurons were collected individually and cellular mRNA was amplified to obtain enough mRNA for sequencing. Sequencing was performed on NextSeq 500, single stranded and 76bp read length.