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. 2014 Feb 10;8:8. doi: 10.3389/fncir.2014.00008

Figure 3.

Figure 3

Morphology of the VPNs labeled by the GAL4 lines with aberrant phototaxis. Right-angled arrows on the bottom corners of the panels indicate directions in the images, A: anterior, L: lateral, D: dorsal. Scale bars correspond to either 20 or 50 μm. (A–D) Three-dimensional (3D) reconstruction of confocal laser scanning sections (horizontal view). Labeled neurons (green-white, visualized with GAL4-driven UAS-GFP) on the background labeling of synaptic neuropils (magenta, with anti-Bruchpilot nc82 antibody). CB indicates labeled cell bodies. In addition to the lobula tangential 11 (LT11) and medulla columnar 61 (MC61) VPNs, NP6099 and MZ820 strains labeled horizontal system (HS) neurons and lobula plate tangential 2 (PT2) neurons, respectively, in the optic lobe. Two arrows in B indicate dark-labeled layers in the background neuropil labeling of the medulla. Box and arrows in C indicate the region and the direction of oblique frontal views shown in Figures 3I, L. ME: medulla, AME: accessory medulla, LO: lobula, LOP: lobula plate, PVLP: posterior ventrolateral protocerebrum, AOT: anterior optic tract, AOTU: anterior optic tubercle, OCH2: second optic chiasmus. (E, F) High-magnification view of the horizontal sections of NP302 and MZ820 strains, showing specific arborizations in the medulla M6 and M7 layers (triangles). Note that the dendrites in the 3D reconstruction images of Figures 3C, D, G, H appear to extend more medially, because they visualize all the arborizations along the curved surface of the M6 and M7 layers. (G, H) Posterior 3D reconstruction of these strains, showing the labeling in different subsets of the MC61 neurons in the dorsal and ventral halves of the medulla. (I, L) 3D reconstruction views of the region shown as a box and arrows in Figure 3C; viewed along the axis that is parallel (I, K) or perpendicular (J, L) to the tangential plane of medulla layers. Entire population of MC61 neurons (I, J) and a sample with two FLP-induced (Wong et al., 2002) single-cell clones (K, L) are shown. In the latter, one of the cell bodies is out of the region of reconstruction (indicated with gray characters). (M, N) Distribution of presynaptic sites (white, visualized with synaptic vesicle-targeting UAS-n-Syb-GFP) and all the neural fibers (red, with cytoplasmic UAS-DsRed). Stacks of confocal horizontal sections. Lo3-5 and M6L, 6M and 7 indicate the layers of dendrites with (white) and without (yellow) presynaptic cites. Note that some of the cell bodies (CBs) are labeled with surplus amount of n-Syb-GFP. (O–P) Labeled cells in the central brain. In addition to the MC61 neurons, NP302 labels glial cells on the surface, dorsal giant interneuron (DGI), mushroom body neurons (MB), terminals of the olfactory sensory neurons (OSN) in the antennal lobe, and putative gustatory and other sensory neurons (GSN) in the gnathal (subesophageal) ganglia. MZ820 labels PT2 neurons as well as extensive glial cells on the surface. See Otsuna and Ito (2006) for the labeling pattern of NP1035 and NP6099. (Q–V) Labeling patterns and phototaxis phenotypes of other GAL4 driver strains that label HS and PT2 neurons, which are labeled simultaneously in the NP6099 and MZ820 strains, respectively, but not neurons of the LT11 or MC61 pathway. Q, R GAL4 strain NP6651 drives expression in the horizontal system cells (HS). The strain also labels the vertical system cells (VS; Note that in total 8–9 VS cells are observed, not 5–7 cells as previously described Heisenberg et al., 1978). S, T GAL4 strain NP1582 drives expression in PT2 neurons. LT13 neurons are also labeled. Reconstruction (horizontal view) of the labeled neurons (green-white, visualized with GAL4-driven UAS-GFP) on the background labeling of autofluorescence (magenta) (Q, S), and posterior views without background labeling (R, T). (U, V) Phototaxis behavior of NP6651 (U) towards 440 nm light and that of NP1582 towards 350- and 570-nm light (V).