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. 2018 Apr 5;7:e34338. doi: 10.7554/eLife.34338

Figure 2. GluRIIA-containing receptors can be knocked down specifically on muscle six using M6 >Gal4.

(A) Schematic of the Type Ib motor neuron innervating both muscles 6 and 7 at the Drosophila NMJ. Gal4 is expressed specifically on muscle six in combination with UAS-GluRIIARNAi using M6 > Gal4 (M6 > GluRIIARNAi: w1118;Tub-FRT-STOP-FRT-Gal4, UAS-FLP, UAS-CD8-GFP/+; H94-Gal4, nSyb-Gal80/UAS-GluRIIARNAi). GluRIIA expression is reduced on muscle 6, while expression of GluRIIA on muscle 7 is unperturbed. (B) Representative images of muscle 6/7 NMJs immunostained with antibodies that recognize the neuronal membrane (HRP) and the postsynaptic GluR subunit GluRIIA in wild type and M6 > GluRIIARNAi. (C) Representative images of individual Ib synaptic boutons immunostained with anti-GluRIIA and anti-GluRIID on the indicated muscles in wild type and M6 > GluRIIARNAi. (D) Quantification of bouton numbers on muscles 6 and 7 in wild type and M6 > GluRIIARNAi. (E) Quantification of mean fluorescence intensity of GluRIIA and GluRIID puncta normalized to wild type on the indicated muscles. Asterisks indicate statistical significance using a Student’s t test: (*) p<0.05; (**) p<0.01; (****) p<0.0001, (ns) not significant. Error bars indicate ± SEM. Detailed statistical information for represented data (mean values, SEM, (n, p) is shown in Supplementary file 1.

Figure 2.

Figure 2—figure supplement 1. Biased Gal-4 expression on muscle six does not alter synaptic growth or function.

Figure 2—figure supplement 1.

(A) Representative images of muscle 6/7 NMJs of wild type and M6 > mCherryRNAi (w1118;Tub-FRT-STOP-FRT-Gal4, UAS-FLP, UAS-CD8-GFP/+; H94-Gal4, nSyb-Gal80/UAS-mCherryRNAi) labeled with phalloidin (actin marker), GFP, and the neuronal membrane marker HRP. (B–D) Quantification of muscle surface area (B), bouton number (C), and BRP puncta number per NMJ (D). No significant differences in any of these parameters were observed. (E) Schematic and electrophysiological traces of recordings from wild type and M6 >mCherryRNAi NMJs. (F–H) Quantification of mEPSP amplitude (F), EPSP amplitude (G), and quantal content (H) from the indicated genotypes finds no significant difference in synaptic physiology. Error bars indicate ± SEM. Asterisks indicate statistical significance using a Student’s t test: (ns) not significant. Detailed statistical information for represented data (mean values, SEM, n, p) is shown in Supplementary file 1.