Figure 6. Ten-a and spectrin function together for normal synapse number.

(A–B) Representative single confocal optical sections taken at equivalent positions of antennal lobes stained for α-spectrin and N-Cadherin in control (A) and ten-a mutant (B) adults. In ten-a mutants, α-spectrin staining is reduced compared to control. The disrupted morphology of the antennal lobe itself is due to partner matching errors evident in the ten-a mutant (Hong et al., 2012). Scale bar = 20 μm. (C–E) Representative confocal Z-stack images of the ORNs in the VA1lm glomerulus expressing Brp-Short in control animals (C), or animals expressing dsRNA against α-spectrin (D) or β-spectrin (E), and stained with antibodies against Brp-Short (red) and N-Cadherin (blue). (F) Quantification of α-spectrin (green), β-spectrin (red), and N-Cadherin (blue) immunofluorescence in control and ten-a mutants. ***p < 0.001. (G) Quantification of Brp-Short puncta in the noted genotypes. In all cases, similar reductions in puncta number are observed. Moreover, the genetic perturbations do not enhance each other, suggesting function in the same pathway. Significance was assessed with a one-way ANOVA and corrected for multiple comparisons by a posthoc Tukey's multiple comparisons test. ***p < 0.001 (compared with control). In all cases, data represent mean ± SEM and n ≥ 10 animals, 19 antennal lobes. Scale bar = 5 μm.

DOI: http://dx.doi.org/10.7554/eLife.03726.017

Figure 6—figure supplement 1. ten-a regulates spectrin levels and spectrin regulates synapse number.

Representative confocal single optical sections of control (A) and ten-a mutant (B) antennal lobes stained with antibodies to β-spectrin (red) and N-Cadherin (blue). Note the reduction (quantified in Figure 6F) of staining in the ten-a mutant. Scale bar = 20 μm. (C–E) Representative high magnification confocal z-stacks of Or67d ORNs expressing Brp-Short-mStraw in control animals (C) and animals co-expressing dsRNA against α-spectrin (D) or β-spectrin (E) and stained with antibodies against mStraw (red) and N-Cadherin (blue). Glomerular morphology is unaffected by spectrin dsRNA but synapse number is reduced. Scale bar = 5 μm. (F) Quantification of Brp-Short puncta in the genotypes from C–E. ***p < 0.0001. In all cases, data represent mean ± SEM and n ≥ 10 animals, 19 antennal lobes.

DOI: http://dx.doi.org/10.7554/eLife.03726.018