Figure 5.

Characterization of the Odd neural projections into the calyx. All images are posterior views of the brain, expect for E, which is an anterior view, and maximum intensity confocal projections. A: Colocalization between Odd neurons (green) and projection neurons (magenta) expressing tdm-tomato in a life brain. Image is a ×40 magnification (zoom 1) composed of eight confocal sections (total depth of 16 μm). All of the Odd neurites within the calyx colocalize with the projection neurons (arrow), showing conclusively that the Odd neurons project into the calyx. B: Colocalization between Odd neurons (magenta) with the OK107-Gal4 line driving GFP (green) to label all Kenyon cells. Image is a ×40 magnification composed of 12 confocal sections (total depth of 24 μm). There is no colocalization between the Odd neurons and OK107-Gal4, showing that the Odd neurons are not part of the Kenyon cell cluster. C: Single-cell MARCM Odd clone (green) induced at 5 hours of embryonic development. Neuropil stained with nc82 (magenta). Image is ×20 magnification composed of 10 confocal sections (total depth of 20 μm). This cell projects a neurite into the calyx (arrow) and IPR. D: Single fluoroform. E: Expression of the dendritic marker DenMark in projection neurons (PNs) driven by GH146-Gal4. Image is a ×20 magnification (zoom 1.5) and is composed of a total number of 18 sections, divided into 12 anterior sections (24 μm thick) and 6 posterior sections (12 μm thick) to better visualize the calic compartment. There are approximately 30 μm between the first 12 sections and the 6 posterior sections. DenMark localizes to the dendritic portion of the PNs (arrows) as shown by DenMark immunostaining visualized with a DsRed antibody (magenta) and colocalization with antennal lobe neurons visualized with nc82 antibody staining (green). There is no antibody localization in the calyx or the lateral horn (arrowheads), indicating that Denmark localizes correctly in PNs. F: Coexpression of the axonal marker synaptotagmin-GFP and the dendritic marker DenMark in the Odd neurons specifically. Dendrites localize to the calyx (∧) whereas the IPR, VLPR, VMPR arbor is pmagentaominantly axonal (*). The projections terminating near the AMMC are also axonal (arrow). Image is a ×20 magnification composed of 15 confocal sections (total depth of 30 μm), showing that the dendritic and axonal markers localize to the different parts of the arbor. G: A high-magnification close-up (×40 magnification with a further zoom of 2) of the calyx labeled with nc82 (green) innervated by the DenMark-labeled portion (magenta) of the Odd projections. Image composed of eight confocal sections (total depth of 16 μm). Scale bar = 50 μm in A–G. [Color figure can be viewed in the online issue, which is available at wileyonlinelibrary.com.]