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. 2020 Sep 2;11:4411. doi: 10.1038/s41467-020-18115-2

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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. 1 — a Schematic of experiment. b Representative images for in vivo recordings of CSF tracer influx after injection in the CM either during the day (orange) or night (blue) under ketamine/xylazine (KX) anesthesia. Dotted white line indicates region of quantification in c. White scale bar: 2 mm. c Mean pixel intensity in arbitrary units (A.U.) over 30 min post CM injection. Thick lines indicate group means with SEM outlined in shaded regions. The inset is the magnitude of fluorescence at 30 min with boxplots, minima is minimum value, maxima is maximum value, center is median and quartiles shown by box and whiskers, individual animals represented as colored dots, two-sided t test, p = 0.0257. Time 0 is tracer infusion start. n = 6 mice per group. Asterisk indicates p < 0.05. d Time course of mean pixel intensity from ex vivo coronal sections under KX (violet, n = ZT22: 10 mice; ZT18: 11 mice; ZT2, ZT6, ZT10, and ZT14: 12 mice), pentobarbital (red, n = ZT2: 7 mice; ZT6, ZT10, and ZT22: 9 mice; ZT18: 10 mice; ZT14: 11 mice), and Avertin (gray, n = ZT22: 7 mice; ZT2 and ZT6: 8 mice; ZT14: 9 mice; ZT10 and ZT18: 10 mice). Each colored point is an animal, black points are mean ± SEM for each time bin. Dotted line indicates estimated cosinor curve. The light cycle is indicated by orange (day) and blue (night) coloring in the background. Zeitgeber Time (ZT) in hours, where ZT0 is lights on and ZT12 is lights off. e Estimated cosine curves aligned at the mesor for KX, pentobarbital (P), and avertin (A), with an inset depicting measurements from a cosine curve. f 95% confidence intervals for estimates of phase, amplitude, and mesor for each anesthetic. Midline is the model generated estimate, bars indicate 95% range around the estimate. Source data are provided as a Source Data file.