Figure 5

Developmental profiling of the glutamatergic ventrolateral cluster. (A) Labelling of developing (third instar larval stage) ventral local interneurons (vILNs) with antibodies against neurotactin (Nrt; magenta). (A1-A3) Sections at 7 μm intervals focus on the cell somata of vlLNs labelled with OK371 (demarcated by white dots). (A1) Neuroblast is not labelled with green fluorescent protein (GFP) (blue arrowhead); GFP co-localizes with neurotactin (Nrt) (white arrowhead), and secondary axonal tract is labelled with anti-Nrt (small white arrows). (A4) Merged image of all 35 confocal sections showing the entire ventrolateral lineage. (B) Reconstruction of appropriate confocal sections using Amira. (B1) Larval antennal lobe (LAL) with ventrolaterally located cell bodies labelled with GFP expression in Gal4-OK371. (B2) Reconstruction of the anti-Nrt labelled lineage; the secondary axonal tract (SAT) traverses the LAL (outlined with dots) and terminates near the prospective adult AL (asterisk). (B3) Merge of the images shown in (B1) and (B2); white dotted line demarcates the location of the neuroblast. (C,D) Antennal lobes from (C) 6 and (D) 12 hours APF, showing developing neurons labelled with Gal4-OK371 (yellow arrowheads). (C2. D2). The presumptive adult antennal lobe is labelled with anti-N-Cadherin (yellow dotted lines). (C3, D3)The regressing larval antennal lobe is identified by staining with mAbnc82 (anti-Brp) (white arrowheads). (D1) Tiled image made from cropped portions of appropriate sections to highlight the cells that are part of this cluster. Orientation of the images; D = dorsal; L = lateral; P = posterior. Scale bar = 10 μm.