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. 2022 Oct 17;11:44. doi: 10.1186/s40035-022-00318-w

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Intestinal overexpression of α-syn recapitulates Parkinson’s disease pathology in Drosophila. a Diagram of the temporal expression of the α-syn transgene in the Drosophila intestine. Temperature-controlled expression of UAS-Syn or UAS-GFP was achieved by co-expression of esg-GAL4 with a temperature-sensitive version of the GAL4 inhibitor Gal80^TS, in which the expression of UAS-synuclein was inactivated and induced at 18 °C and 29 °C, respectively. We refer to the two genotypes as esg^TS > Syn (w; esg-Gal4, UAS-GFP/+; UAS-Syn/tubulin-Gal80^TS) and esg^TS > GFP (w; esg-Gal4, UAS-GFP/+; UAS-GFP/tubulin-Gal80^TS). b Western blot showing the expression of human α-syn in the midgut of adult flies after 48 h at 29 °C. α-Actin was used as a loading control. c Immunostaining showing the expression of human α-syn in fly intestinal stem cells (Scale bar, 1 mm). α-Syn expression in intestinal stem cells increased following a shift to 29 °C. d Representative images showing exogenous α-syn in both cytoplasm and nucleus (Scale bar, 1.5 μm). Red: α-syn; green: GFP; blue: DNA. e Decreased lifespan of intestinal PD flies. Adult flies were cultured as described in panel a and the percentage of living male flies was calculated to monitor lifespan. f Survival rates of intestinal PD flies challenged with rotenone. Sucrose solution containing 7.5 mM rotenone was used for survival assay. g Intestinal overexpression of α-syn causes an age-dependent dopaminergic (DA) neuron loss. Numbers of TH-positive DA neurons in five clusters of each genotype were counted and analyzed (bottom). Representative images of TH-positive DA neurons in the PPM1 cluster of 28-day-old control or intestinal α-syn-expressing flies (top). Scale bars, 0.2 μm. h Expression of α-syn in the midgut leads to progressive locomotor deficits. For the locomotion assay, climbing distances of each genotype of males at different time points are plotted as bar graphs. Mean ± SEM; P-values of survival curves were calculated using log-rank tests (using total fly numbers), and for category graphs using one-way ANOVA with a Bonferroni multiple-comparison test. *P < 0.05; **P < 0.01; ***P < 0.001