Skip to main content
. 2017 May 8;7(7):2023–2038. doi: 10.1534/g3.117.041632

Figure 4.

Figure 4

Neuronal depletion of candidate essential chaperones alter NMJ morphology in Drosophila. (A–J) Confocal images of NMJ synapses at muscle 6/7 of (A) control, elav-Gal4 driven (B) CG4183 RNAi, (C) CG15266 RNAi, (D) CG7770 RNAi, (E) CG7048 RNAi, (F) CG17187 RNAi, (G) CG5525 RNAi, (H) CG12101 RNAi, (I) CG8542 RNAi, and (J) CG1242 RNAi double immunolabeled with a presynaptic marker (CSP, red) and neuronal membrane marker (HRP, green) to reveal the bouton outline at the NMJs. Compared to the control NMJ, elav-Gal4-mediated depletion of the above-mentioned essential chaperones showed significantly altered NMJ morphology. The Roman numerals in the image correlate the NMJ morphological defect of each panel with the corresponding phenotypic class depicted in Figure 3. The bouton numbers were determined by manually counting the number of CSP-positive varicosities at the NMJs. Bar in (J) represents 20 µm. (K) Histogram showing average number of boutons at muscle 6/7 of A2 hemisegment in the control, elav-Gal4/+, CG4183 RNAi, elav-Gal4-driven CG4183 RNAi, CG15266 RNAi, elav-Gal4-driven CG15266 RNAi, CG7770 RNAi, elav-Gal4-driven CG7770 RNAi, CG7048 RNAi, elav-Gal4-driven CG7048 RNAi, CG17187 RNAi, elav-Gal4-driven CG17187 RNAi, CG5525 RNAi, elav-Gal4-driven CG5525 RNAi, CG12101 RNAi, elav-Gal4-driven CG12101 RNAi, CG8542 RNAi, elav-Gal4-driven CG8542 RNAi, CG1242 RNAi, and elav-Gal4-driven CG1242 RNAi. Histogram in black represents controls including w1118, elav-Gal4/+, and the parental lines. Total number of boutons upon pan-neuronal knockdown of sHSP (green), Prefoldins (magenta), Hsp40 (violet), Hsp60 (orange), Hsp70 (dark blue), and Hsp90 (light blue) have been represented. At least eight NMJ synapses of A2 hemisegments from four larvae of each genotype were used for bouton quantification. ** P < 0.001 and *** P < 0.0001. Error bars represent SEM (mean ± SEM). Statistical analysis based on one-way ANOVA with post hoc Tukey’s test for multiple comparisons. CSP, cysteine string protein; HRP, horseradish peroxidase; Hsp, heat shock protein; NMJ, neuromuscular junction; ns, not significant; RNAi, RNA interference; sHSP, small HSP.