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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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The heterozygous Ace mutation fully suppresses the ChAT/+ ’s inhibitory control deficits. (A) The double heterozygous ChAT^MI04508/Ace^c00215 (light purple) mutants displayed robust movement restraint with rarely detectable LIEs similar to CS (blue), Ace^c00215/+ (light green) and ChAT^MI04508/+ (orange) at the age of 4 day (one way ANOVA with post hoc Tukey: ns, p > 0.05; n = 12). At the ages of 2 week and 4 week, the ChAT^MI04508/Ace^c00215 mutants displayed a small increase in LIE similar to CS and Ace^c00215/+ but the level was significantly different from that of ChAT^MI04508/+ (ANOVA with post hoc Tukey: ***, p < 0.0001; n = 12). (B) The latency to the initial LIE was comparable in all genotypes under study at the age of 4 day (ANOVA with post hoc Tukey: ns, p > 0.05; n = 12). The double heterozygous ChAT^MI04508/Ace^c00215 exhibited the significantly delayed latency compared to that of ChAT^MI04508/+ at the ages of 2 week and 4 week (ANOVA with post hoc Tukey: different letters denote statistically significant difference, p < 0.01, n = 12). (C) The acetylcholine levels in the double heterozygous ChAT^MI04508/Ace^c00215 were comparable to CS, but significantly lower than Ace^c00215/+ and significantly higher than ChAT^MI04508/+ at all ages. ANOVA with post hoc Tukey multiple comparison of the genotypes within an age: ns, p > 0.05; **, p < 0.005; ***, p < 0.0001; n = 4–8.