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. Author manuscript; available in PMC: 2010 Feb 12.
Published in final edited form as: Neuron. 2009 Feb 12;61(3):373–384. doi: 10.1016/j.neuron.2008.12.033

Figure 8. E49 motor neurons do not synapse on gustatory sensory neurons.

Figure 8

(A) Expression of CD2::GFP under the control of Gr5a-LexA::VP16 detected by immunofluorescence.

(B,C) Single labeled E49 motor neurons (magenta) and projections from Gr5a (B) or Gr66a (C) gustatory neurons (green). Genotypes are tub>Gal80>/Gr5a-LexA::VP16; E49-Gal4, UAS-CD8::dsRed; lexAop-CD2::GFP/MKRS, hsFLP and tub>Gal80>/Gr66a-Ires-GFP; E49-Gal4/UAS-CD8::dsRed; MKRS, hsFLP. Dashed box in (B) indicates size and approximate position of images presented in H and I.

(D) GRASP between Gr5a neurons expressing CD4::spGFP11 and GAD1 neurons expressing CD4::spGFP1-10 and CD8:dsRed (magenta). GRASP detected by immunofluorescence using monoclonal GFP antibody (green; Sigma). Genotype is GAD1-Gal4/lexAop-CD4::spGFP11; Gr5a-LexA::VP16/UAS-CD4::spGFP1-10. Dashed box indicates approximate position of images in E and F.

(E,F) GRASP between Gr5a neurons expressing CD4::spGFP11 and 2 GAD1 neurons expressing CD4:spGFP1-10. GRASP detected by immunofluorescence using monoclonal GFP antibody (green; Sigma). GAD1 cells expressing CD4::spGFP1-10 identified using rabbit polyclonal GFP antibody (magenta; Abcam). Genotype is Gr5a-LexA::VP16/tub>Gal80>; LexAop-CD4::spGFP11/GAD1-Gal4; UAS-CD4::spGFP1-10/MKRS, hsFLP.

(G) Expression of CD4::spGFP1-10 under control of E49-Gal4 detected by immunofluorescence against GFP1-10 (Abcam polyclonal antibody).

(H,I) Lack of GRASP observed between E49 neurons co-expressing CD4::spGFP1-10 and CD8::dsRed (magenta), and Gr5a neurons expressing CD4::spGFP11. Panels are representative images of E49 dendrites in Gr5a region. GRASP detected by immunofluorescence using polyclonal GFP antibody (green; Invitrogen). Arrow in (I) indicates infrequently observed GRASP signal. Dashes outline approximate position of Gr5a cells in each panel. Genotype is Gr5a-LexA::VP16/+; LexAop-CD4::spGFP11/E49-Gal4; UAS-CD4::spGFP1-10/UAS-CD8::dsRed.

(J–L) Lack of GRASP between Gr5a neurons expressing CD4::spGFP11 and a single labeled E49 motor neuron co-expressing CD4::spGFP1-10 and CD8::dsRed (magenta). GRASP detected by immunofluorescence using monoclonal GFP antibody (green; Sigma). (K) and (L) are enlargements of indicated areas from (J). Arrows in (L) show infrequently observed GRASP signals. Dashes outline approximate position of Gr5a cells in (K). Genotype is Gr5a-LexA::VP16/tub>Gal80>; LexAop-CD4::spGFP11/E49-Gal4, UAS-CD8::dsRed; UAS-CD4::spGFP1-10/MKRS, hsFLP.

Scale bar is 50 μm in (A) and applies to all panels not noted otherwise. Scale bars in (E.F) are 20 μm and (H,I,K,L) are 10 μm. “E49” indicates expression in all E49-Gal4 neurons, “E49 MN” indicates expression in a single E49 motor neuron.