Establishment of Tools for Neurogenetic Analysis of Sexual Behavior in the Silkmoth, Bombyx mori
Figure 1
Visualization of cellular surface by mCD8GFP expression in silkmoths.
(A–F) GFP fused with the membrane-tethering signal polypeptide (mCD8GFP) was ubiquitously expressed by crossing UAS-mCD8GFP and Actin A3-GAL4 strains. GFP fluorescent images of the brains of second instar larva (A) and male adult moth (B), and pupal body (C, D). Brightfield images (B′–D′). Red color in the eyes is due to DeRed expression by the 3xP3 promoter used as a selection marker (B′, C′). L: Left brain hemisphere, R: Right brain hemisphere. (E, F) GFP expression was visualized with antibody staining (green). Nuclear DNA was visualized with DAPI (blue). Pictures of maximum intensity projection (E and F) or a single optical plane (E′ and F′) of confocal images of the whole brain (E and E′) and the antennal lobe (F and F′). Note that the GFP signal is detected at the cellular surface. (G, H) Visualization of sex pheromone receptor neurons by crossing UAS-mCD8GFP and BmOR1-GAL4 strains. (G) GFP and glomerular structure of the antennal lobe was visualized with anti-GFP and anti-synapsin antibody, respectively. Each neural projection was visible (arrowhead) and axon terminals were correctly targeted to the bombykol-responsive glomeruli (toroid of the macroglomerular complex [MGC]). Gs: Ordinary glomeruli. (H) Each receptor cell (arrowhead) and neurite (arrow) was visible. Scale bars: 100 µm (A, E–H), 1 mm (B–D).