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Loss of bmm in the neurons with an independent RNAi line has no effect on triglyceride storage or breakdown in females.

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posted on 14.04.2021, 16:23 by Lianna W. Wat, Charlotte Chao, Rachael Bartlett, Justin L. Buchanan, Jason W. Millington, Hui Ju Chih, Zahid S. Chowdhury, Puja Biswas, Vivian Huang, Leah J. Shin, Lin Chuan Wang, Marie-Pierre L. Gauthier, Maria C. Barone, Kristi L. Montooth, Michael A. Welte, Elizabeth J. Rideout

(A) Whole-body triglyceride storage in 5-day-old virgin females overexpressing UAS-bmm-RNAi in the postmitotic neurons (elav>UAS-bmm-RNAi) was not significantly different from age-matched control females (elav>+ and +>UAS-bmm-RNAi) (p = 0.54 and 0.95, respectively; one-way ANOVA followed by Tukey HSD test). (B) There was a significant reduction in whole-body triglyceride levels in 5-day-old elav>+, +>UAS-bmm-RNAi, and elav>UAS-bmm-RNAi females between 0 and 12 hours STV (p = 0.026, 0.0038, and 0.0013, respectively; one-way ANOVA followed by Tukey HSD test). Between 12 and 24 hours STV, 5-day-old elav>+, +>UAS-bmm-RNAi, and elav>UAS-bmm-RNAi females modestly decreased triglyceride levels by similar magnitudes (p = 1.2 × 10−4, 0.15, and 0.0012, respectively; one-way ANOVA followed by Tukey HSD test). (C) Whole-body triglyceride storage in females with elav-GAL4-mediated overexpression of an additional UAS-bmm-RNAi#2 (VDRC #37877) transgene in neurons was not significantly different from control females (elav-GAL4>+ and +>UAS-bmm-RNAi#2) (p = 0.56 and 0.035, respectively; one-way ANOVA followed by Tukey HSD test). (D) There was a modest decrease in triglyceride levels between 0 and 12 hours and 12 and 24 hours STV among elav-GAL4>UAS-bmm-RNAi#2 (VDRC #37877) females and elav-GAL4>+ and +>UAS-bmm-RNAi#2 controls (p = 0.011, 0.0027, and 1.4 × 10−6 for 0–12 hours STV and 4.6 × 10−4, 3.4 × 10−4, and 0.002 for 12–24 hours STV, respectively; one-way ANOVA followed by Tukey HSD test). (E) Whole-body triglyceride levels in 5-day-old virgin nSyb>UAS-bmm-RNAi females were not significantly different to nSyb-GAL4>+ and +>UAS-bmm-RNAi controls (p = 0.85 and 0.52, respectively; one-way ANOVA followed by Tukey HSD test). (F) Triglyceride breakdown post-starvation was modestly decreased by a similar magnitude among genotypes in 5-day-old virgin nSyb>UAS-bmm-RNAi females and control females (nSyb>+ and +>UAS-bmm-RNAi) between 0 and 12 hours STV (p = 0.21, 0.017, and 0.43, respectively; one-way ANOVA followed by Tukey HSD test). Between 12 and 24 hours STV, the decrease in triglyceride levels in nSyb>UAS-bmm-RNAi females was blocked, whereas triglyceride levels decreased in control nSyb-GAL4>+ and +>UAS-bmm-RNAi females during this interval (p = 0.093, 3.0 × 10−5, and 0.0019, respectively; one-way ANOVA followed by Tukey HSD test). (G) Whole-body triglyceride levels in 5-day-old virgin elav>UAS-bmm;UAS-bmm-RNAi females were not significantly different to elav-GAL4>+ and +>UAS-bmm;UAS-bmm-RNAi controls (p = 0.41 and 0.66, respectively; one-way ANOVA followed by Tukey HSD test). (H) Whole-body triglyceride levels post-starvation among 5-day-old virgin elav>UAS-bmm;UAS-bmm-RNAi females and control females (elav>+ and +>UAS-bmm;UAS-bmm-RNAi) decreased by a similar magnitude between 0 and 12 hours or 12 and 24 hours STV, demonstrating that re-expression of UAS-bmm rescued the effects of bmm loss in neurons STV (p = 0.0023, 0.0012, and 0.0027 for 0–12 hours and 1.0 × 10−6, 5.2 × 10−6, and 2.9 × 10−4 for 12–24 hours, respectively; one-way ANOVA followed by Tukey HSD test). Asterisks indicate a significant difference between two sexes, two genotypes, or two time points (*p < 0.05, **p < 0.01, ***p < 0.001). Error bars on graphs depicting percent body fat represent SEM; error bars on graphs depicting the change in percent body fat represent COE. See S1 Table for a list of all multiple comparisons and p-values; quantitative measurements underlying all graphs are available in S1 Data. bmm, brummer; COE, coefficient of error; elav, embryonic lethal abnormal vision; F, female; HSD, honest significant difference; ns, no significant difference between two sexes, two genotypes, or time points; nSyb, neuronal Synaptobrevin; STV, post-starvation; UAS, upstream activation sequence; VDRC, Vienna Drosophila Resource Center.

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