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. 2019 Mar 26;218(5):1706–1724. doi: 10.1083/jcb.201807165

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© 2019 Goel et al.

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Figure 6. — Manipulating presynaptic cargo transport to NMJ terminals by arl8 modulates the gain of synaptic strength at endo and rab3 NMJs. (A) Representative images of type Ib NMJ boutons immunostained with antibodies that recognize AZ components (BRP, Unc13A), neuronal membrane (HRP), and SV markers (vGlut, Synaptotagmin [SYT], and Synapsin [SYN]) in wild type, arl8 mutants (arl8: w;arl8^e00336), and neuronal overexpression of arl8 (arl8-OE: w;OK6-Gal4,UAS-arl8-GFP/+). Note that the number of AZs and the intensity of SV markers at NMJ terminals are reduced by loss of arl8 (C; Table S1), while arl8-OE enhances the intensity of AZ and SV markers (C; Table S1). (B) EPSP traces in the indicated genotypes demonstrate that synaptic strength is reduced by loss of arl8 and increased by arl8-OE. (C) Quantification of the indicated values in the three genotypes normalized to wild-type values (n ≥ 12; one-way ANOVA; Table S1). (D–F) Representative images (D), EPSP traces (E), and quantification (F) for NMJs of endo^RNAi and endo^RNAi combined with arl8-OE (endo^RNAi+arl8-OE: w;OK6-Gal4,UAS-arl8-GFP/+;UAS-endo-RNAi). arl8-OE enhances synaptic strength and BRP and vGlut intensity above endo^RNAi baseline values (note that values are normalized to endo^RNAi; n ≥ 7; Student’s t test; Table S1). (G–I) Representative images (G), EPSP traces (H), and quantification (I) for NMJs of rab3^RNAi, rab3, and arl8 double mutants (rab3;arl8: w;rab3^rup;arl8^e00336), and rab3^RNAi combined with arl8-OE (rab3^RNAi+arl8-OE: w;OK6-Gal4,UAS-arl8-GFP/+;UAS-rab3-RNAi). Note that loss of arl8 reduces BRP puncta number and intensities of BRP and vGlut below rab3^RNAi baseline values, while arl8-OE enhances these values. Values are normalized to rab3^RNAi (n ≥ 9; one-way ANOVA; Table S1). Error bars indicate ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.