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. 2021 Feb 9;11:3432. doi: 10.1038/s41598-021-82813-0

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Dop1R1 signaling in the mushroom body mediates ethanol and methamphetamine preference. (a) Dop1R1 is necessary for acquired ethanol preference. Daily preference for 15% ethanol is plotted for the wild type (CS) and mutant strains for the four dopamine receptors. One-way repeated-measures ANOVA or the Friedman test is performed among the indices from the day 1 to 4. CS: P < 0.0001, n = 24 (Friedman); Dop1R1: P = 0.2004, n = 22, (Friedman); Dop1R2: F _{(2.176, 43.52)} = 6.715, P = 0.0023, n = 21 (ANOVA); Dop2R: F _{(1.676, 38.55)} = 5.037, P = 0.0155, n = 24 (ANOVA); DopEcR: F _{(2.021, 40.42)} = 5.731, P = 0.0063, n = 21 (ANOVA). (b) Dop1R1 expression in the mushroom body is necessary for acquired ethanol preference. Dop1R1 is genetically knocked down using the MB010B-GAL4 driver. MB010B/CS: F _{(1.719, 56.73)} = 4.093, P = 0.0271, n = 34 (ANOVA); MB010B/UAS-Dop1R1.RNAi: F _{(2.806, 78.57)} = 2.282, P = 0.0896, n = 29 (ANOVA); w/UAS-Dop1R1.RNAi: F _{(1.979, 43.54)} = 6.227, P = 0.0043, n = 23 (ANOVA). (c) Neurotransmission from the PAM cluster dopamine neurons is necessary for acquired ethanol preference. PAM cluster neurons are blocked throughout the measurement using the R58E02-GAL4 and the UAS-shibire^ts1 strains and the 15% ethanol preference is plotted. R58E02-GAL4/w: F _{(2.663, 47.93)} = 9.746, P < 0.0001, n = 19 (ANOVA); R58E02-GAL4/UAS-shi^ts1: F _{(2.382, 52.41)} = 1.718, P = 0.1840, n = 23 (ANOVA); w/UAS-shi^ts1: F _{(2.705, 119.0)} = 6.435, P = 0.0007, n = 45 (ANOVA). Note that female flies were used for the measurement because of the high mortality when males were used (see also Supplementary Figure 1). (d) Dop1R1 is necessary for acute methamphetamine preference. Daily preference for 0.3 mM methamphetamine is plotted for the wild type (CS) and mutant strains for the four dopamine receptors. The drug preference of mutant strains is compared to that of the wild type on the first day (Dunnett’s multiple comparison). CS vs Dop1R1: P < 0.0001, CS vs Dop1R2: P = 0.0052, CS vs Dop2R: P = 0.8207, CS vs DopEcR: P = 0.2005. (e) Dop1R1 expression in the mushroom body is necessary for acute methamphetamine preference. Dop1R1 is genetically knocked down using the MB010B-GAL4 driver. The drug preference on the first day is compared among genotypes (Dunn’s multiple comparison). MB010B-GAL4/CS vs. MB10B-GAL4/UAS-DopR1.RNAi: P = 0.0193; MB010B-GAL4/UAS-DopR1/RNAi vs. w/UAS-DopR1.RNAi: P = 0.0001. Bar graphs: mean ± SEM. *: P < 0.05.