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. 2019 May 2;9:6811. doi: 10.1038/s41598-019-43080-2

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© The Author(s) 2019

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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Ventral midbrain dopaminergic neurons show intrinsic electrical properties different from ventral midbrain non-dopaminergic neurons. (A) Live images in bright field (top picture) and the corresponding mOrange and Fluo-4 AM (middle and bottom pictures) fluorescent signal of differentiated neurons (Day 50) during calcium imaging. Squares in the bottom picture point to mOrange⁺ dopaminergic neurons whereas circles point to mOrange⁻ non-dopaminergic neurons in which oscillations of Fluo-4 AM fluorescence are measured. (B) Calcium traces of selected mOrange⁺ and mOrange⁻ neurons confirm electrophysiological activity. Blue arrowheads point to neuronal firing events. Insets show in detail calcium traces from individual mOrange⁺ or mOrange⁻ neurons. Red line represents an increase in fluorescence equivalent to 3 standard deviations (SD) of the mean of the baseline noise. (C) Quantification of the average amplitude of the firing events of mOrange⁺ and mOrange⁻ after 35 and 50 days of differentiation. Data show the average ± S.E.M. of two or three independent experiments. At day 35, 130 mOrange⁺ and 1187 mOrange⁻ cells from 3 independent experiments. At day 50, 124 mOrange⁺ and 1034 mOrange⁻ cells from 2 independent experiments. Asterisk denotes statistically significant differences (*p < 0.05).