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. 2022 Dec 3;12:20903. doi: 10.1038/s41598-022-25402-z

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© The Author(s) 2022

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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Aging diminishes inhibitory control in flies. (A) Representative movement traces of a group of 13 CS flies subjected to the GNG test in three chambers—one with the 4 day old, the other with the 2 week old and the last with the 4 week old. Each panel illustrates individual fly traces, which are denoted by the different colored lines. The timepoint when strong airflow was introduced is marked as 0 in the X axis. The CS flies of all three ages exhibited robust inhibitory control under airflow with infrequent instances of flying behavior representing LIE (movement > 60 mm/s). The sporadic LIEs increased with aging. (B) Quantitative representation of the aging-dependent increases in LIE and rescue by Ace deficiency. All LIEs displayed by 13 flies in a chamber for the entire 10 min under airflow during GNG test were manually counted revealing more frequent LIE in older CS flies (blue line). The aging-dependent increases in the LIE were alleviated by the heterozygous acetylcholinesterase (Ace) mutation (green line; Ace^c00215/+) fully at the 2 week old and partly at the 4 week old. ANOVA with post hoc Tukey multiple comparison of three ages in each genotype: different letters (A, B and C for CS; a and b for Ace^c00215/+) denote statistically significant differences (p < 0.05; n = 12). Student t-test between two genotypes (Ace^c00215/+ and CS) in each age: ns, p > 0.05; ***, p < 0.0001; n = 12. (C) Latency to initial LIE. ANOVA with post hoc Tukey multiple comparison of three ages in each genotype: different letters (A and B for CS) denote statistically significant differences (p < 0.0001; n = 12). When the age-matching CS and Ace^c00215/+ were compared by Student t-test, there is no difference in the LIE latency at 4 day or 2 week (p > 0.05, n = 12; not noted in the graph) but there is significant difference at 4 week (**, p < 0.005, n = 12). The heterozygous Ace mutation fully reversed the aging-related premature LIE latency. (D) Acetylcholine levels in the CS and Ace^c00215/+ heads. The acetylcholine levels were significantly higher in Ace^c00215/+ than CS at all ages and dampened with aging in both genotypes. ANOVA with post hoc Tukey multiple comparison of three ages in each genotype: different letters (A, B and C for CS; a, b and c for Ace^c00215/+) denote statistically significant differences (p < 0.05; n = 4). Student t-test between two genotypes (Ace^c00215/+ and CS) in each age: **, p < 0.005; n = 4.