PHP and sustained memory consolidation depend on Myo15. (A) Confocal images of Myo15-RNAi–mediated knock-down and control NMJs treated with PhTx to induce PHP stained for BRP (fire LUT), scale bar 2 µm. (B) Quantification of A, BRP mean pixel intensity (control - PhTx 100.0 ± 7.21%, n = 12; + PhTx 142.5 ± 5.67%, n = 11; Myo15-RNAi - PhTx 100.0 ± 8.54%, n = 15; + PhTx 97.16 ± 5.65%, n = 11). (C) Representative traces of evoked EPSPs after 10 min PhTx treatment at control (black) and Myo15-RNAi knock-down (red) third-instar muscle 6/7 NMJs, scale bars 10 ms, 10 mV. (D and E) Quantification of 10 min PhTx treatment, n = 10. (D) Normalized mEPSPs (control - PhTx 1.00 ± 0.066%; + PhTx 0.46 ± 0.021%; Myo15-RNAi - PhTx 1.00 ± 0.056%; + PhTx 0.58 ± 0.038%). (E) Normalized eEPSPs (control - PhTx 1.00 ± 0.067%; + PhTx 0.96 ± 0.076%; Myo15-RNAi - PhTx 1.00 ± 0.077%; + PhTx 0.98 ± 0.045%). (F) Normalized quantal content (control - PhTx 1.00 ± 0.096%; + PhTx 2.03 ± 0.176%; Myo15-RNAi - PhTx 1.00 ± 0.088%; + PhTx 1.71 ± 0.134%). (G) Representative traces of evoked EPSPs after 30 min PhTx treatment at control (black) and Myo15-RNAi knock-down (red) third-instar muscle 6/7 NMJs, scale bars 10 ms, 5 mV. (H–J) Quantification of 30 min PhTx treatment, n = 10. (H) Normalized mEPSPs (control - PhTx 1.00 ± 0.072%; + PhTx 0.636 ± 0.028%; Myo15-RNAi - PhTx 1.00 ± 0.034%; + PhTx 0.57 ± 0.031%). (I) Normalized eEPSPs (control - PhTx 1.00 ± 0.114%; + PhTx 1.02 ± 0.076%; Myo15-RNAi - PhTx 1.00 ± 0.048%; + PhTx 0.48 ± 0.073%). (J) Normalized quantal content (control - PhTx 1.00 ± 0.121%; + PhTx 1.59 ± 0.176%; Myo15-RNAi - PhTx 1.00 ± 0.055%; + PhTx 0.77 ± 0.115%). (K) Schematic representation of aversive olfactory learning setup. (L) Quantification of MTM 1 h (control driver line PI 0.21 ± 0.028, Myo15 knock-down (RNAi101729) PI 0.08 ± 0.033, control RNAi PI 0.22 ± 0.026, F(2,23) = 7.056, P = 0.0045, n = 8, post hoc Tukey’s multiple comparisons test, Gal80ts;ok107-Gal4/+ versus Gal80ts;ok107-Gal4/RNAi104604, *P < 0.05, +/RNAi104604 versus Gal80ts;ok107-Gal4/RNAi104604, **P < 0.01). (M) Quantification of STM (control driver line PI 0.27 ± 0.057, Myo15 knock-down (RNAi101729) PI 0.24 ± 0.031, control RNAi PI 0.27 ± 0.038, F(2,17) = 0.1446, P = 0.8666, n = 6). All data are provided as mean ± SEM. (B–J)n = NMJs with 1–2 NMJs/animal, four to six animals, L and M: n = cohort of ∼50 adult flies, 6–8 cohorts measured. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05; n.s., not significant. PI, performance index.
Figure 7.

PHP and sustained memory consolidation depend on Myo15. (A) Confocal images of Myo15-RNAi–mediated knock-down and control NMJs treated with PhTx to induce PHP stained for BRP (fire LUT), scale bar 2 µm. (B) Quantification of A, BRP mean pixel intensity (control - PhTx 100.0 ± 7.21%, n = 12; + PhTx 142.5 ± 5.67%, n = 11; Myo15-RNAi - PhTx 100.0 ± 8.54%, n = 15; + PhTx 97.16 ± 5.65%, n = 11). (C) Representative traces of evoked EPSPs after 10 min PhTx treatment at control (black) and Myo15-RNAi knock-down (red) third-instar muscle 6/7 NMJs, scale bars 10 ms, 10 mV. (D and E) Quantification of 10 min PhTx treatment, n = 10. (D) Normalized mEPSPs (control - PhTx 1.00 ± 0.066%; + PhTx 0.46 ± 0.021%; Myo15-RNAi - PhTx 1.00 ± 0.056%; + PhTx 0.58 ± 0.038%). (E) Normalized eEPSPs (control - PhTx 1.00 ± 0.067%; + PhTx 0.96 ± 0.076%; Myo15-RNAi - PhTx 1.00 ± 0.077%; + PhTx 0.98 ± 0.045%). (F) Normalized quantal content (control - PhTx 1.00 ± 0.096%; + PhTx 2.03 ± 0.176%; Myo15-RNAi - PhTx 1.00 ± 0.088%; + PhTx 1.71 ± 0.134%). (G) Representative traces of evoked EPSPs after 30 min PhTx treatment at control (black) and Myo15-RNAi knock-down (red) third-instar muscle 6/7 NMJs, scale bars 10 ms, 5 mV. (H–J) Quantification of 30 min PhTx treatment, n = 10. (H) Normalized mEPSPs (control - PhTx 1.00 ± 0.072%; + PhTx 0.636 ± 0.028%; Myo15-RNAi - PhTx 1.00 ± 0.034%; + PhTx 0.57 ± 0.031%). (I) Normalized eEPSPs (control - PhTx 1.00 ± 0.114%; + PhTx 1.02 ± 0.076%; Myo15-RNAi - PhTx 1.00 ± 0.048%; + PhTx 0.48 ± 0.073%). (J) Normalized quantal content (control - PhTx 1.00 ± 0.121%; + PhTx 1.59 ± 0.176%; Myo15-RNAi - PhTx 1.00 ± 0.055%; + PhTx 0.77 ± 0.115%). (K) Schematic representation of aversive olfactory learning setup. (L) Quantification of MTM 1 h (control driver line PI 0.21 ± 0.028, Myo15 knock-down (RNAi101729) PI 0.08 ± 0.033, control RNAi PI 0.22 ± 0.026, F(2,23) = 7.056, P = 0.0045, n = 8, post hoc Tukey’s multiple comparisons test, Gal80ts;ok107-Gal4/+ versus Gal80ts;ok107-Gal4/RNAi104604, *P < 0.05, +/RNAi104604 versus Gal80ts;ok107-Gal4/RNAi104604, **P < 0.01). (M) Quantification of STM (control driver line PI 0.27 ± 0.057, Myo15 knock-down (RNAi101729) PI 0.24 ± 0.031, control RNAi PI 0.27 ± 0.038, F(2,17) = 0.1446, P = 0.8666, n = 6). All data are provided as mean ± SEM. (B–J)n = NMJs with 1–2 NMJs/animal, four to six animals, L and M: n = cohort of ∼50 adult flies, 6–8 cohorts measured. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05; n.s., not significant. PI, performance index.

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