Figure 3. Neurotransmitter expression and co-expression in the larval neuronal population.

(A) Neurons are classified based on the expression of neurotransmitters. Seurat UMAP plot showing the distribution of the five main neuronal cell-types. (B) Simplified heatmap, representing neuronal subpopulations. Genes displayed are the main markers analyzed to identify neuronal cell-identities. The x axis represents individual cells, each line corresponds to one neuron. Gene expression levels are color coded. (C) Neurotransmitters are co-expressed in a subset of neurons. Dot size corresponds to the percentage of cells expressing a particular gene, while color intensity represents gene expression levels. (D) UpSet plot (Conway et al., 2017) illustrating the co-expression of neurotransmitters. Light and bold numbers represent number of cells and percentages of cells, respectively. The percentages were calculated based on positive cells for a particular neurotransmitter or a combination of them, in comparison to the total of neurons from the dataset. (E) Co-expression analysis based on the simultaneous expression of ple and VGlut. Blend UMAP plot showing only ple positive cells. Cells co-expressing ple and VGlut are shown in yellow. Threshold: 0,4.

Figure 3—figure supplement 1. Neurotransmitter co-expression in the larval brain.

(A) Neurotransmitters are co-expressed with monoamines. VGlut and ple overlapping. (B–C) Neurotransmitters co-expression. Subsets of Glutamatergic/GABAergic neurons and GABAergic/Cholinergic neurons. Neurons with the overlap of interest are circled with a dashed-line and pointed with arrows. Nuclei were labeled with DAPI. Scale bar: 10 µm.

Figure 3—figure supplement 2. Monoaminergic neurons analysis.

(A) Re-clustering analysis of monoaminergic neurons revealed an enrichment of Dopaminergic neurons. UMAP plot indicating the expression of dopaminergic marker genes: Vmat, ple and Ddc, against other aminergic markers: Tbh, Tdc2 and SerT. (B) The remaining aminergic neurons can be identified in different clusters in the original UMAP plot, validating the co-expression between neurotransmitters. Gene expression levels are color coded.