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. 2020 Jul 28;11:3764. doi: 10.1038/s41467-020-17543-4

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© The Author(s) 2020

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. 5 — a Representative coronal ×4 optical plane through the VTA showing fluorescence indicative of mCherry (red; top; hM4Di reporter) and TH (green; bottom; Alexa 488), and corresponding merged image (right). b The number of cells (n = 948 cells across four rats) counted in each 300 × 300 μm optical section. c Cell counts indicate the specificity of mCherry expression in TH+ neurons and the proportion of TH+ neurons that were transfected (n = 948 cells across four rats). d Representative area of a ×40 image used for the analyses in (b) and (c). Cells with colocalized TH and mCherry signals all had a nucleus indicated by DAPI (blue). e Example traces show averaged excitatory post synaptic currents (EPSCs) for an NAc core medium spiny neuron (MSN) before (baseline), during (+CNO), and after (washout) 5 min application of 1 μM CNO to striatal slices containing hm4Di-expressing terminals from VTA dopamine neurons. f Peak amplitudes of EPSCs recorded from individual MSNs innervated by hM4Di-expressing dopaminergic terminals (n = 8 cells across five rats). g Normalized mean EPSC amplitudes for the same group of cells. Clozapine-n-oxide reduced EPSC amplitude to 65.7 ± 7.3% of baseline [n = 8 cells across five rats; F_(2,13) = 6.77, p = 0.01; Newman−Keuls p = 0.01]. Mean EPSC amplitude returned towards baseline values within 20 min of washout (93.9 ± 8.9% of baseline). h Example traces show averaged EPSCs for an MSN before (baseline), during (+CNO), and after (washout) 10 min application of CNO to striatal slices containing excitatory hM3Dq-expressing terminals from VTA dopamine neurons. i Peak amplitudes of EPSCs recorded from individual MSNs innervated by hm3Dq-expressing terminals (n = 9 cells across five rats). j Normalized mean EPSC amplitudes for the same group of cells. Clozapine-n-oxide significantly increased EPSCs to 137.9 ± 18.8% of baseline levels, [n = 9 cells across five rats, F_(2,14) = 17.84, p < 0.001; Newman−Keuls p < 0.001]. Responses returned to baseline values within 20 min of washout (102.9 ± 4.7% of baseline). Averaged data are mean ± s.e.m. Data were analysed using RM ANOVA (g, j) and Newman−Keuls post hoc tests (g, j). All statistical tests were two-sided. Raw data are available as a supplementary source data file.