RNA profile of nuclei isolated from neurons coexpressing VAChT and VGlut or VAChT and VGAT

The concept of neurotransmitter identity, once considered singular and immutable for mature neurons, has been broadened by the recognition that many neurons release multiple neuro-active substances. In order to demonstrate the extent of neurotransmitter co-expression, we labelled neuronal nuclei with UNC84-GFP using VAChT and either Vglut or VGAT split-GAL4s and isolated them with the isolation of nuclei tagged in specific ell types (INTACT) method From the isolated nuclei, we isolated and sequenced Poly(A) RNA and examined the relative expression levels of glutamatergic, GABAergic, or cholinergic genes in each population. Comparative gene expression profiling analysis of RNA-Seq data from INTACT purified nuclei from VAChT:Vglut split Gal4/UAS-UNC84-GFP and VAChT:VGAT split Gal4/UAS-UNC84-GFP flies