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. 2017 Dec 12;6:e32027. doi: 10.7554/eLife.32027

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© 2017, Huang et al

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Figure 5. — (a) Diagram of Drosophila circadian neurons. The two groups of PDF LNvs are located close to optic lobe: four l-LNvs (black) have dendrites that branch into the ipsilateral optic lobe, and project their axons across the central brain to the contralateral optic lobe. Four s-LNvs (red) project their dorsal axons to dorsoposterior part of the central brain, where DN1 (blue), DN2 (purple) and DN3 (green) neurons are located. (b) Labeling of neurons that receive synaptic input from PDF cells expressing no ligand (top panels: b1, (b3, b5, b7), or the ligand nSyb::CD19 driven by the pdf-LexA driver (bottom panels: b2, (b4, b6, b8). To unbiasedly identify downstream synaptic targets of PDF neurons, we used the pan-neuronal receptor strain, nSybE-nlgSNTG4. b1, b2: nSyb::CD19 +expression pattern. b1: control brain with no nSyb::CD19 expression. b2: nSyb::CD19 driven by pdf-LexA accumulated (red) in the cell bodies (yellow arrows) and the axon terminals of s-LNv dorsal axons (yellow arrowheads). b3: No GFP induction was observed in the control brains without pdf >nSyb::CD19 ligand. b4: Induction of CD4::tdGFP expression in pdf >nSyb::CD19 brain in the vicinity of the trajectory of nSyb::CD19 +axons through the central brain. Arrow and arrowhead point to GFP +neurons in the DN1 region, and DN3 regions, respectively. b5, b6: Immunostaining with anti-PER antibody identifies DN1 (white arrow), DN2, and DN3 (white arrowhead) neurons. b7, b8: Merged images of PER (magenta), GFP (green) and nSyb::CD19 (red). Stippled square in b8 indicates region shown at high magnification (40X) in Figure 6.

Figure 5—figure supplement 1. — (a) Diagram of Drosophila circadian neurons. The two groups of PDF LNvs are located close to optic lobe: four l-LNvs (black) have dendrites that branch into the ipsilateral optic lobe, and project their axons across the central brain to the contralateral optic lobe. Four s-LNvs (red) project their dorsal axons to dorsoposterior part of the central brain, where DN1 (blue), DN2 (purple) and DN3 (green) neurons are located. (b) Labeling of neurons that receive synaptic input from PDF cells expressing no ligand (top panels: b1, (b3, b5, b7), or the ligand nSyb::CD19 driven by the pdf-LexA driver (bottom panels: b2, (b4, b6, b8). To unbiasedly identify downstream synaptic targets of PDF neurons, we used the pan-neuronal receptor strain, nSybE-nlgSNTG4. b1, b2: nSyb::CD19 +expression pattern. b1: control brain with no nSyb::CD19 expression. b2: nSyb::CD19 driven by pdf-LexA accumulated (red) in the cell bodies (yellow arrows) and the axon terminals of s-LNv dorsal axons (yellow arrowheads). b3: No GFP induction was observed in the control brains without pdf >nSyb::CD19 ligand. b4: Induction of CD4::tdGFP expression in pdf >nSyb::CD19 brain in the vicinity of the trajectory of nSyb::CD19 +axons through the central brain. Arrow and arrowhead point to GFP +neurons in the DN1 region, and DN3 regions, respectively. b5, b6: Immunostaining with anti-PER antibody identifies DN1 (white arrow), DN2, and DN3 (white arrowhead) neurons. b7, b8: Merged images of PER (magenta), GFP (green) and nSyb::CD19 (red). Stippled square in b8 indicates region shown at high magnification (40X) in Figure 6.