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. Author manuscript; available in PMC: 2010 Dec 10.
Published in final edited form as: Neuron. 2009 Dec 10;64(5):663–677. doi: 10.1016/j.neuron.2009.11.002

Figure 2. The rup active zone phenotype is due to loss of rab3 function.

Figure 2

(A) Immunostaining with an antibody against Rab3 (green) shows the distribution of Rab3 protein at WT NMJs and the complete loss of wild type Rab3 protein at rup mutant NMJs. Partial concentration of Rab3 at active zones is observed by co-staining with anti-Brp (red). Inset shows a magnified image of boxed region. Scale bars represent 2 μm in lower magnification images and 1 μm in higher magnification inset.

(B) The Rab3 antibody recognizes a single band on an immunoblot from wild type larval brain extracts that is missing in protein extracts prepared from rup mutant larval brains. Tubulin band serves as a loading control.

(C, D, and E) Rescue of the rup active zone phenotype by expression of a UAS-rab3 transgene driven in neurons by ELAVGeneSwitch. (C) NMJs from WT (ELAVGeneSwitch/+), rup mutant (rup; ELAVGeneSwitch/+), and rup mutant larvae expressing UAS-rab3 (rup; ELAVGeneSwitch/UAS-rab3) were immunostained with α-Brp (red) and α-DGluRIII (green) to visualize the apposition of Brp puncta with DGluRIII clusters and Brp puncta area. The larvae of all three genotypes were maintained on food containing RU486. Scale bar represents 2 μm. NMJs from the genotypes listed in (C) were quantified for the percentage of DGluRIII clusters apposed to Brp puncta per NMJ (D) and the average area of individual Brp punctum (E). n = 10 NMJs for all genotypes; *p≪0.001 vs. wild type and rescue.