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Fig. 3. Confocal images of 40 µm sections showing RFamide immunoreactivity
(green, Cy2) during AL development. (A) Area of the larval antennal center
(LAC) in a wandering larva (W2, 2-day-old wandering stage shortly before
entering the prepupal stage). Within the developing lateral cell group (LC,
encircled area) type II (arrow) and III (arrowheads) somata are stained. The
LAC is labeled (red) by dextran backfill of the antennal nerve. The double
arrowhead marks a dextran-labeled axon bundle coming from the larval antenna.
(B–F) Double immunolabeling with the RFamide antiserum (green, Cy2) and
the synaptotagmin antibody (red, Cy5). (B) Anterior section through the
developing AL showing the type V neuron entering the AL from the tritocerebrum
(TC). (C) In stage P9 developing glomeruli (GL) can be distinguished by
synaptotagmin immunostaining. RFamide immunostaining in basal parts of the
developing glomeruli is weak compared to staining of the type V arborizations
(arrowheads) and to staining intensity in stages later than P12/13 (see
D–E). In the LC the large type II cell (large arrow) with its neurite
leaving the AL (smaller arrows) and smaller type I cells are labeled. (D)
Frontal section through posterior parts of a stage P16 AL with strongly
labeled type I cells, strong labeling in the glomeruli (GL) and intensely
labeled fibers in the root of the outer antenno-cerebral tract (arrows). (E)
Oblique section showing typical labeling of glomeruli in an adult AL. The type
V cell enters the AL neuropil from the TC (arrow). (F) Magnification from E
showing the neurite of the type V cell entering the AL neuropil. A–E,
frontal sections; orientation bars in C (D, dorsal; M, medial) also apply to
A–D; CN, coarse neuropil; Es, Esophagus; MC, median cell group; MGC,
macroglomerular complex. Scale bars, 40 µm (A,B); 80 µm (C–E); 20
µm (F).
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