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. 2014 Apr 9;8:35. doi: 10.3389/fncir.2014.00035

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Copyright © 2014 Masuda-Nakagawa, Ito, Awasaki and O'Kane.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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A single larval GABAergic neuron, resembling the adult APL neuron, innervates the larval calyx. (A) NP0732-GAL4 expression, visualized using UAS-mCD8::GFP, is seen in two large cell bodies, in a stereo image of an anterior view of a 3D reconstruction of the larval central brain. One of these (asterisk) posterior to the MB sends a neurite laterally, which branches upwards to the MB calyx (Ca) and downwards to arborizations (labeled in the GFP channel) in (1) the terminal regions of the medial lobe (ML), (2) the proximal region of the vertical lobe (VL), and (3) an area in the region connecting the lower pedunculus (Pe) and vertical lobe. The MB is labeled using anti-Dlg. (B) A projection of four confocal sections through the medial lobe of the same preparation. GABA is found in the NP0732-GAL4 neuron (arrowheads), although weakly and sporadically in its fine arborizations, as well as in one or more neurons innervating the lobes that are not labeled by NP0732-GAL4 (arrows) (C) Confocal section of a larval calyx, showing numerous GABAergic termini, all labeled also with mCD8::GFP expressed under control of NP0732-GAL4. Some large GABAergic termini (arrowheads) lie between calyx glomeruli (labeled with anti-Dlg). The two surface plots in the bottom row represent the intensities in the GABA and mCD8::GFP channels; note that virtually all GABA peaks lie within regions of mCD8::GFP expression in the larval APL. (D) A similar confocal section and plots of a larval calyx showing overlap of GABAergic termini with mCD8::GFP expressed under control of NP2631-GAL4. (E) A similar confocal section of a larval calyx showing overlap between GABAergic termini, and axonal and synaptic termini (examples shown with arrowheads) expressing mCD8::GFP under control of GH146-GAL4, which is also expressed in a subset of PNs (examples of calyx glomeruli containing these are shown with asterisks). (F) Octopaminergic (OA) termini in the calyx do not overlap with termini of the larval APL (examples shown with arrowheads) labeled with mCD8::GFP expressed under control of NP2631-GAL4. Scale bars 10μm.