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. 2019 May 21;8:e43079. doi: 10.7554/eLife.43079

Figure 2. The most specific split-GAL4 lines for identified cholinergic LHONs.

Array of projections of the most specific split-GAL4 line for each cholinergic LHON cell-type. Where available, a neuron from that class traced to at least to identification in the EM volume (inset, black represents tracing, grey the lateral horn, blue and brown the single neuron labelling, Nern et al., 2015). Expression of split-GAL4 lines are visualized using UAS-csChrimson::mVenus in attP18 (green), with nc82 as a neuropil stain (magenta). Each image is registered to JFRC2010, while the EM data is registered to JFRC2013. The cell-type (eg. PV5a1) labelled is in the top left of each panel, while the line code (eg. LH290) is listed below. Cell-types that stained positive for more than one neurotransmitter are labelled with an asterix in the top right. Off-target expression in the brain for non-ideal lines labelled with a yellow arrow. See www.janelia.org/split-gal4 for image data.

Figure 2.

Figure 2—figure supplement 1. Examples of cell type staining positive for two neurotransmitters, examples of bilateral LHONs.

Figure 2—figure supplement 1.

(A’) Projection of half a brain with cell type AV7a1 labelled. The grey box represents the axons of the neuron where we focused our imaging for the neurotransmitter staining of acetylcholine and glutamate. Note: these neurons did not stain positive for GABA (data not shown). (A’’) Single confocal slice showing colabelling of the axonal boutons of AV7a1 (labelled with csChrimson::mVenus in green) with a stain for ChAT in magenta (marker for cholinergic neurons). Blue arrows label two particular presynaptic sites where colocalization is visible indicating these cells are cholinergic. (A’’’) Single confocal slice showing colabelling of the axonal boutons of AV7a1 (labelled with csChrimson::mVenus in green) with a stain for VGlut in grey (marker for glutamatergic neurons). Blue arrows label two particular presynaptic sites (same sites as in A’’) where colocalization is visible indicating these cells are also glutamatergic. (B’) Projection of a confocal stack of a single bilateral cholinergic AD3b1 neuron using MultiColor FlpOut (MCFO) in green, registered to the JFRC2013 template brain. (B’’) Projection of a confocal stack of a single bilateral cholinergic AD1e1 neuron using MultiColor FlpOut (MCFO) in green, registered to the JFRC2013 template brain. (B’’’) Projection of a confocal stack of a single bilateral glutamatergic PV5k1 neuron using MultiColor FlpOut (MCFO) in green, registered to the JFRC2013 template brain.