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. 2020 Oct 20;9:e62953. doi: 10.7554/eLife.62953

Figure 5. FlpTag, a new tool for cell-type-specific, endogenous labeling as shown with GluClα.

Figure 5.

(A) Scheme of FlpTag cassette (first panel) and integration of FlpTag cassette into target gene (second panel). The FlpTag cassette consists of attB-sites, specific FRT sites which form a FLEx-switch, a splice acceptor, GFP and a splice donor. After ΦC31-dependent integration of the FlpTag cassette into a coding intron of the GluClα target gene, two lines with opposite orientations of the cassette can be obtained. In the initial line with the cassette and GFP in opposite orientation with respect to the gene (shown here), the cassette is spliced out together with the intron and no GFP-labeling occurs. After cell-type-specific Flp expression, the FlpTag cassette is flipped, stably integrated as an artificial exon and GluClα is labeled with GFP. (B) Optic lobe with T4/T5 neurons labeled with myr::tdTomato and FlpTag-GluClα::GFP. Left panel: horizontal view on the optic lobe overview (scale bar: 20 μm). Central panel: close-up of medulla layer M10, lobula layer Lo1 and Lobula plate layers 1–4 (scale bar: 5 μm). Right panel: Frontal view on medulla layer M10 with T4 dendrites (scale bar: 20 μm); inset: close-up of columnar GluClα::GFP structure in layer 10 of the medulla. (C) Close-up of FlpTag-GluClα::GFP driven with a T4-Gal4-line; shown are layer 10 of the medulla where T4 dendrites reside and lobula plate layers 1–4 where T4 project their axon terminals to (scale bar: 5 μm). (D) Close-up of FlpTag-GluClα::GFP driven with a T5-Gal4-line; shown are layer 10 of the medulla where T4 dendrites reside and lobula plate layers 1–4 where T4 project their axon terminals to (scale bar: 5 μm).