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. 2021 May 5;11:9579. doi: 10.1038/s41598-021-88910-4

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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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Nicotine promotes dopaminergic neuron survival. A) Dopaminergic neuron loss over time in control (UAS-GFP/+; th-GAL4/+) and experimental (UAS-GFP/UAS-Sph; th-GAL4/+) genotypes; untreated populations are represented by solid lines and treated populations by dotted lines. No significant differences were evidenced between the treated and untreated control genotype. Flies expressing Sph-1 in dopaminergic neurons have significantly fewer neurons than the control genotype throughout their life. However this difference becomes smaller between control and the Sph- expressing treated population as flies age. B) The number of surviving TH neurons in each experimental condition at the last time point studied (Day 60) evidences a reduction in th-positive neurons in the Sph-1 expressing flies compared with control genotype (#### p < 0.0001). Interestingly, this analysis also shows that nicotine treatment does not affect the number of th-positive neurons in the control genotype (p > 0.05), but it does so in the Sph-1 expressing flies (# p < 0.05 compared with the control genotype not treated with nicotine). The Sph-1 expressing flies treated with nicotine show a partial recovery in the number of th-positive neurons, since it is significantly increased compared with not-treated Sph-1 expressing flies (*p < 0.05), but does not reach the number of positive neurons recorded in the control genotype (*p < 0.05). In each case, data from controls were represented in black bars; data from animals treated with nicotine were represented in red bars. All statistical analysis were from two-way ANOVA followed by Tuckey’s post-hoc test, n = 5 fly brains. In all cases experimental animals were treated with 24 µM nicotine.