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. 2022 Mar 31;11:e71469. doi: 10.7554/eLife.71469

Figure 2. Expression of Lar and Sns reporters in motor neurons.

(A–D) Confocal projections of 4–6 optical slices showing EGFP expression driven by either LarMI02154-T2A-GAL4 (Lar>GFP) or SnsMI03001-T2A-GAL4 (Sns>GFP) (green) co-stained with anti-HRP (blue). The bright paired midline cells include motor neurons (A, C, arrows). (E–H’) Confocal projections of larval neuromuscular junctions (NMJs) on muscles 7/6, 13, and 12 (E, F’) and zoomed-in on muscle 7/6 (G, H’), triple-stained with anti-GFP (green), anti-HRP (red), and anti-Dlg (blue). (E’, F’, G’, H’) show GFP signal only. Anti-HRP labels neuronal membranes, and anti-Dlg labels the subsynaptic reticulum at 1b boutons. Lar>GFP and Sns>GFP expression is seen in both 1b and 1s boutons (green), while only Sns>GFP is seen in type II boutons (F’, arrows). (I, J) Projection of optical slices through an entire larval hemisegment showing Lar>GFP (I) and Sns>GFP (J) expression in both 1b and 1s motor neurons. Individual muscles are numbered. Dorsal is to the right. Note that Lar>GFP is equally expressed in most axons and NMJs, while Sns>GFP is expressed at lower levels in axons and NMJs of motor neurons projecting to dorsal muscles. (K, L) Close-up of NMJs on muscles 3 and 4 showing both Lar>GFP (K) and Sns>GFP (L) expression in 1b and 1s NMJs on those muscles. Scale bar, 20 µm. See Figure 2—figure supplement 1 for further characterization of Lar and Sns expression in the larval ventral nerve cord (VNC) and central nervous system (CNS).

Figure 2.

Figure 2—figure supplement 1. Lar and Sns reporter expression in the larval ventral nerve cord (VNC) and brain.

Figure 2—figure supplement 1.

(A–A’’) Sns expression visualized by EGFP reporter expression driven by SnsMI03001-T2A-GAL4 (Sns>GFP; green), co-stained with Even-skipped antibody (Eve) in the larval VNC. Projection of four optical slices is shown. Sns is expressed in Eve-positive RP2/MNISN-1s neurons. (B, B’) Dendritic projections of Sns-expressing neurons seen by driving Drep2 (dendrite-specific marker) with Sns-GAL4 (green), co-stained with FasII antibody to label the mushroom body neuropil (blue). Dendritic projections of Sns-expressing neurons are seen enveloping the dorsal lobe of the mushroom body (B’, arrow). (C–D’) Sns is not expressed in glial cells. (C) Projection of confocal slices through the mushroom body showing no Sns co-expression in Repo-positive glial cells (green). Sns is visualized by driving mCD8-RFP reporter expression using Sns-GAL4 (red). (D, D’) Projection of confocal slices through the entire larval brain, showing no Sns expression in glial cells. Single slice is shown in D’. Scale bar, 20 µm.