Fig. 1: Chronic striatum-wide measurements of rapid ACh release dynamics in head-fixed, behaving mice. | Nature Communications

Fig. 1: Chronic striatum-wide measurements of rapid ACh release dynamics in head-fixed, behaving mice.

From: Distinct spatially organized striatum-wide acetylcholine dynamics for the learning and extinction of Pavlovian associations

Fig. 1

a Schematic of the fiber array approach for measuring ACh release across the striatum. b Spherical treadmill and imaging set up for head-fixed mice. c ACh3.0 fluorescence traces (∆F/F) measured from 14 example fibers in the striatum of a single mouse. d Left: Post-mortem micro-CT scan images in the coronal plane (top) and axial plane (bottom) from a representative mouse. Fibers appear in white. Right: Images from the Allen Brain Common Coordinate Framework Atlas corresponding to the CT planes on left. Red circles indicate the position of an automatically localized fiber tip. e Locations of all fibers used for ACh measurements in the axial (left) and sagittal (right) planes. Each dot is the location (relative to bregma) of a single fiber and the color indicates the mouse identity (n = 8 mice). f Top: mean ∆F/F aligned to unpredicted water reward delivery for a single fiber from a single mouse for single sessions (n = 8 reward deliveries/session) at 1–3 weeks from the first day of Pavlovian training (Fig. 2). Inset indicates fiber location in the coronal plane. Shaded regions, S.E.M. Bottom: The mean peak (early/late) or dip ∆F/F for unpredicted reward deliveries across 18 consecutive sessions of training (n = 8 reward deliveries/session) for the example at top. Values are the mean ± S.E.M. For each component, the mean of each day was compared to the mean of every other day using one-way ANOVA, p = 0.1371 and F(17,129) = 1.420 for early peak, p = 0.1751 and F(17,129) = 1.346 for dip, p = 0.9897 and F(17,129) = 0.367 for late peak. g Top: percentage of the total fibers (295 fibers) across all mice (n = 8) with significant (p < 0.01, two-tailed Wilcoxon rank-sum test, see “Methods”) component(s) to unpredicted reward delivery. Bottom: maps as in (e) showing the presence of each combination of signal components to unpredicted reward delivery for each fiber. Empty circles indicate no significant response. h Top: Mean ∆F/F aligned to unpredicted reward delivery with quasi-simultaneous 405 nm (black) and 470 nm (blue) illumination in a mouse expressing the non-functional mutant ACh3.0 sensor. Shaded regions, S.E.M. Bottom: Histogram of Pearson’s correlation coefficients between ∆F/F fluorescence traces obtained with quasi-simultaneous 405 nm and 470 nm illumination in mutant ACh sensor expressing mice (n = 3). i Mean unpredicted reward triggered ∆F/F from a single fiber in functional ACh3.0 sensor (top) and mutant ACh sensor (bottom) expressing mice before and after 0.3 Hz high pass filtering. Shaded region, S.E.M. Note that the small, slow artifactual decrease in the mutant sensor recording is largely eliminated with filtering, but the rapid reward triggered release measured with the functional sensor is preserved. Brain schematic in (a), mouse schematic in (b), objective in (b), and water drop schematics in (f), (h) and (i) were adapted from SciDraw (scidraw.io) and are licensed under CC BY 4.0 (https://creativecommons.org/licenses/by/4.0/). Source (https://doi.org/10.5281/zenodo.3925911, https://doi.org/10.5281/zenodo.3925913, https://doi.org/10.5281/zenodo.4914800, and https://doi.org/10.5281/zenodo.3925935), respectively. Brain schematics in (e) and (f) were adapted from the Allen Mouse Brain Common Coordinate Framework (CCFv3) (https://atlas.brain-map.org/). Source data are provided as a Source data file.

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