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. 2020 Jun 29;21(13):4601. doi: 10.3390/ijms21134601

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Locomotor activity is altered in nlg3^Def1 flies. (A,B) Startle-induced climbing represented by the mean percent ± s.e.m. of flies reaching the top vial. (A) nlg3^Def1 males had less flies than Cs reaching the top vial at 3–4 days old (two-way ANOVA –effect of genotype: F_1,31 = 4.464, p = 0.0428). However, this was not the case at 7–10 days old, where there was no difference between Cs and nlg3^Def1. Cs males also had less flies reaching the top vial at 7–10 days old compared to 3–4 days old but this difference was not observed in nlg3^Def1 (two-way ANOVA—effect of age F_1,31 = 9.391, p = 0.0045; interaction between age and genotype F_1,31 = 4.780, p = 0.0365). (B) nlg3^Def1 females had less flies than Cs reaching the top vial at both ages (two-way ANOVA—effect of genotype: F_1,30 = 8.238, p = 0.0075); however flies at 7–10 days old had a lower percentage reaching the top vial compared to 3–4 day-old flies (two-way ANOVA—effect of age: F_1,30 = 23.38, p < 0.0001). n = 15 trials of 40 flies for all treatments. (C) elav > nlg3 males had no difference in the number of flies reaching the top vial at 3–4 days old compared to the genotype controlled elav-Gal4/+ and UAS-nlg3/+ (two-way ANOVA—effect of genotype: F_2,38 = 1.809, p = 0.1776). At 7–10 days old there was a decrease in the number of elav-Gal4/+ and UAS-nlg3/+ flies reaching the top vial (two-way ANOVA—effect of age: F_1,38 = 29.39, p < 0.0001). However, elav > nlg3 males at 7–10 days old did not show this reduction in percentage of flies reaching the top vial (two-way ANOVA—interaction of age and genotype: F_2,38 = 3.993, p = 0.0267). (D) elav > nlg3 females mirror this trend. The 3–4 day-old females also had no difference in the number of flies reaching the top compared to the genotype controls of the same age (two-way ANOVA—effect of genotype: F_{2, 36} = 0.09254, p = 0.9118). As well, elav-Gal4/+ and UAS-nlg3/+ 7–10 day-old females decrease in the number of flies reaching the top vial (two-way ANOVA—effect of age: F_{1, 36} = 42.12, p < 0.0001). The 7–10 day-old elav > nlg3 females also had a reduction in the age-related decline of startle-induced climbing although not statistically significant (two-way ANOVA—interaction of age and genotype: F_2,36 = 1.760, p = 0.1865). C,D: n = 5–9 trials of 40 flies. (E,F) Spontaneous locomotor activity presented as violin plots of the mean number of beam crossings. (E) Age and genotype affect locomotor activity in males (Kruskal–Wallis: p = 0.0004), and male nlg3^Def1 had increased locomotor activity compared to Cs at 7–10 days old but not at 3–4 days old (Dunn’s post hoc: p = 0.0032). (F) In females a similar trend was observed (Kruskal–Wallis: p = 0.0367), where nlg3^Def1 7–10 days old had higher activity than Cs (Dunn’s post hoc: p = 0.0806). E,F: n = 15 individual flies. Dotted horizontal lines represents 25–75% quartiles, solid horizontal line displays the median. Those are not visible when 75% of the data = 0. Lighter colors represent 3–4 day-old flies. Darker colors represent 7–10 day-old flies.

Locomotor activity is altered in nlg3^Def1 flies. (A,B) Startle-induced climbing represented by the mean percent ± s.e.m. of flies reaching the top vial. (A) nlg3^Def1 males had less flies than Cs reaching the top vial at 3–4 days old (two-way ANOVA –effect of genotype: F_1,31 = 4.464, p = 0.0428). However, this was not the case at 7–10 days old, where there was no difference between Cs and nlg3^Def1. Cs males also had less flies reaching the top vial at 7–10 days old compared to 3–4 days old but this difference was not observed in nlg3^Def1 (two-way ANOVA—effect of age F_1,31 = 9.391, p = 0.0045; interaction between age and genotype F_1,31 = 4.780, p = 0.0365). (B) nlg3^Def1 females had less flies than Cs reaching the top vial at both ages (two-way ANOVA—effect of genotype: F_1,30 = 8.238, p = 0.0075); however flies at 7–10 days old had a lower percentage reaching the top vial compared to 3–4 day-old flies (two-way ANOVA—effect of age: F_1,30 = 23.38, p < 0.0001). n = 15 trials of 40 flies for all treatments. (C) elav > nlg3 males had no difference in the number of flies reaching the top vial at 3–4 days old compared to the genotype controlled elav-Gal4/+ and UAS-nlg3/+ (two-way ANOVA—effect of genotype: F_2,38 = 1.809, p = 0.1776). At 7–10 days old there was a decrease in the number of elav-Gal4/+ and UAS-nlg3/+ flies reaching the top vial (two-way ANOVA—effect of age: F_1,38 = 29.39, p < 0.0001). However, elav > nlg3 males at 7–10 days old did not show this reduction in percentage of flies reaching the top vial (two-way ANOVA—interaction of age and genotype: F_2,38 = 3.993, p = 0.0267). (D) elav > nlg3 females mirror this trend. The 3–4 day-old females also had no difference in the number of flies reaching the top compared to the genotype controls of the same age (two-way ANOVA—effect of genotype: F_{2, 36} = 0.09254, p = 0.9118). As well, elav-Gal4/+ and UAS-nlg3/+ 7–10 day-old females decrease in the number of flies reaching the top vial (two-way ANOVA—effect of age: F_{1, 36} = 42.12, p < 0.0001). The 7–10 day-old elav > nlg3 females also had a reduction in the age-related decline of startle-induced climbing although not statistically significant (two-way ANOVA—interaction of age and genotype: F_2,36 = 1.760, p = 0.1865). C,D: n = 5–9 trials of 40 flies. (E,F) Spontaneous locomotor activity presented as violin plots of the mean number of beam crossings. (E) Age and genotype affect locomotor activity in males (Kruskal–Wallis: p = 0.0004), and male nlg3^Def1 had increased locomotor activity compared to Cs at 7–10 days old but not at 3–4 days old (Dunn’s post hoc: p = 0.0032). (F) In females a similar trend was observed (Kruskal–Wallis: p = 0.0367), where nlg3^Def1 7–10 days old had higher activity than Cs (Dunn’s post hoc: p = 0.0806). E,F: n = 15 individual flies. Dotted horizontal lines represents 25–75% quartiles, solid horizontal line displays the median. Those are not visible when 75% of the data = 0. Lighter colors represent 3–4 day-old flies. Darker colors represent 7–10 day-old flies.