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. 2021 Aug 18;7:76. doi: 10.1038/s41531-021-00210-w

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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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a (Top) Representative spontaneous calcium activity in naive dopaminergic neurons (left, black) and dopaminergic neurons overexpressing α-syn (right, pink) exemplify the alteration in calcium dynamics due to increased levels of α-syn. (Bottom) Spontaneous calcium activity encompassing all neurons recorded in each experimental group (n = 33 wild-type neurons, n = 40 α-syn-overexpressing neurons, form eight biological replicates). b Calcium events in all neurons. c Spontaneous calcium event rate, width, and amplitude. Overexpression of α-syn does not alter calcium event rate (two-tailed unpaired t test, WT vs. α-syn, p = 0.1775, n = 33 wild-type neurons, n = 40 α-syn-overexpressing neurons), α-syn burden broaden calcium events (p = 0.0152, two-tailed unpaired t test, WT vs. α-syn, n = 33 wild-type neurons, n = 40 α-syn-overexpressing neurons) and increases in amplitude (p = 0.0000198, two-tailed unpaired t test, WT vs. α-syn, n = 33 wild-type neurons, n = 40 α-syn-overexpressing neurons). d Representative whole-cell current-clamp recordings of spontaneously active naive (top left, black) compared with overexpressing α-syn dopaminergic neurons (top right, pink). Distribution of raw interspike intervals (ISIs) in naive (bottom left) and α-syn-overexpressing neuron (bottom right) (raw ISI distribution, Kolmogorov–Smirnov test, D = 0.26529, p < 0.001). e Naive compared with α-syn-overexpressing dopaminergic neurons (firing frequency: from eight independent experiments, 100 ± 21.21 for naive neurons vs. 281.7 ± 61.30 for α-syn-overexpressing neurons, two-tailed unpaired t test, p = 0.0142; f interspike interval (ISI): 100 ± 21.63 for naive neurons compared to 43.55 ± 13.28 for α-syn-overexpressing neurons, two-tailed unpaired t test, p = 0.0431) and g firing regularity trend in bursts with intermediated periods of quiescence (CV of ISI—100 ± 18.59 for naive neurons vs. 174.9 ± 35.20 for α-syn-overexpressing neurons, two-tailed unpaired t test, p = 0.0808). Empty circles in panel c represent statistical outliers included in the analyses. Bar graphs ± SEM are overlaid with individually filled data points. *p < 0.05; ****p < 0.0001.