Full brains (A–C) or one antennal lobe (D–I) were imaged from flies of the following genotypes: +/+; GH146-Gal4/LexAop-CD4:spGFP11; Or83b-LexA::VP16/UAS-CD4::spGFP1-10 (A,D,G), +/+; GH146-Gal4/LexAop-CD4::spGFP11; +/UAS-CD4::spGFP1-10 (B,E,H) and +/+; +/LexAop-CD4::spGFP11; Or83b-LexA::VP16/UAS-CD4::spGFP1-10 (C,F,I). Or83b-LexA::VP16 drives expression in 80% of olfactory sensory neurons, and GH146-Gal4 drives expression in second order projection neurons that project dendrites to about 30 glomeruli (Jefferis et al., 2007; Lai et al., 2008; Marin et al., 2002).
(A–C) Unfixed brains were imaged for GFP fluorescence. Long dashes outline brain and short dashes outline antennal lobes.
(D–F) The antennal lobes of fixed brains were imaged for immunofluorescence against GFP to detect GRASP (green; Invitrogen rabbit polyclonal antibody) and the neuropil (magenta; nc82). Arrows indicate weak detection of CD4::spGFP1-10 in PN cell bodies; this polyclonal antibody weakly recognized CD4::spGFP1-10 expressed alone.
(G–H) The antennal lobes of fixed brains were imaged for immunofluorescence against GFP to detect GRASP (green; mouse monoclonal antibody). No signal was observed in the absence of GFP reconstitution. Dashes outline antennal lobe. Scale bars are 50 μm.