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. 2021 Mar 31;12:1994. doi: 10.1038/s41467-021-22260-7

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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. 4 — a Injection and fiber placement protocol. b Representative example of GCaMP6m-expression and fiber placement. c Expression and fiber placement at AP + 2.76. Shaded areas: GCaMP6-expression. Stars: fiber tips. d Example traces from GCaMP6m-expressing and eYFP-expressing animals. Dots represent trialstarts. e Variable delay trials from one rat, by the trial outcome. Left two plots: 5 s delay trials, synchronized on trialstart (left) or response (right). Middle: 7.5 s delay. Right: 12.5 s delay. δF/F z-scored to trial baseline (5 s–1 s before trialstart). f Same as e, color-coded by the outcome. Dots represent frames with δF/F > 2std above baseline. Colors same as in d. g Average correct trial fluorescence. h Area under the curve (AUC) during the correct trial delay. Delay, short vs. long, p_MDL = 0.0002; p_MDM = 0.014; p_DMS = 0.0006; p_VMS = 0.0016. i Fluorescence rise kinetics 1 s after trialstart, a fraction of peak fluorescence. Correct, p_MDL-MDM < 0.0001; p_DMS-VMS < 0.0001, p_MDL-VMS = 0.0095; Omissions, p_MDL-MDM = 0.002, p_MDL-VMS = 0.0029; Premature, p_MDL-MDM = 0.0006, p_MDL-VMS = 0.0001, p_DMS-MDM = 0.0017, p_DMS-VMS = 0.0002. j Time to the first synchronous event. Correct, p_MDL-VMS = 0.0007, p_DMS-VMS = 0.006; Omissions, p_MDL-VMS = 0.0344, p_DMS-VMS = 0.0344; Premature p_MDL-VMS = 0.0102, p_DMS-VMS = 0.002. k Total time signal > baseline + 2std. Correct p_MDL-VMS = 0.0093, p_DMS-VMS = 0.0241; Omissions p_MDL-MDM = 0.0285, p_MDL-VMS = 0.0127, p_DMS-VMS = 0.0285; Premature p_MDL-VMS = 0.0008, p_MDL-VMS = 0.0383, p_DMS-VMS = 0.0008. l Peak fluorescence during the delay. Correct, p_MDL-VMS = 0.0215, p_DMS-VMS = 0.0493; Omissions, p_MDL-MDM = 0.0448, p_MDL-VMS = 0.034, p_DMS-VMS = 0.034; Premature, p_MDL-VMS = 0.0294, p_DMS-VMS = 0.0078. Dots represent individual animals in h–l. Shades in g represent SEM. Scale in e is the same for all heatplots. Boxplots in h–l: center line, median; edges, 1st and 3rd quartile; whiskers, range. Friedman with Dunn’s used in h. Kruskal–Wallis with uncorrected Dunn’s and FDR used in i–l. Significance: *p < 0.05. Scales: (a) 400 μm, (d) horizontal 50 s, vertical 10% δF/F, (g): horizontal 2 s, vertical 1% δF/F. Group sizes: MDL (n = 8 rats), MDM (n = 4), DMS (n = 7), VMS (n = 6).