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. 2018 Apr 18;24(8):1206–1219. doi: 10.1038/s41380-018-0034-4

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© The Author(s) 2018

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 — Activation of CB1R and group I mGluR, obligatory components of endocannabinoid system in CA3-CA1, is similar between Pyr-WT and Pyr-KO. a Fluorescent micrographs of CA1 St. radiatum from Pyr-WT (left) and Pyr-KO (right) showing VGlut1 (red) and CB1R immunostaining (green). Overlap of the two signals is yellow. No apparent difference is detectable between the two genotypes. Calibration bar: 5 µm. b Western Blot analysis for CB1R, relative to GAPDH, for Pyr-WT and Pyr-KO. Left—image of a representative gel showing CB1R specific and GAPDH specific bands. Right—bar graph of relative CB1R protein abundance in Pyr-WT (1.26 ± 0.07, n = 5) vs. Pyr-KO (1.24 ± 0.04, n = 5), p = 1.000, U-test. c Cartoon of CA3-CA1 synapse depicting effect of CB1R agonist WIN-55,212-2 [1] on CB1R activation, resulting in lower glutamate release [2]. d 15 min bath application of 5 µm WIN-55,212-2 generates persistent WIN-LTD in Pyr-WT (49.4 ± 0.95 %; n = 8 slices/4 mice; p < 0.001 relative to baseline) and Pyr-KO (65.9 ± 0.89 %; n = 7 slices/3 mice; p < 0.001 relative to baseline). e Average PPR for WIN-LTD in Pyr-WT: 1.59 ± 0.11 (n = 8 slices/4 mice) vs. Pyr-KO: 1.34 ± 0.10 (n = 7 slices/3 mice); p = 0.10. f Cartoon depicting effect of Group I mGluR agonist DHPG [1] on eCB release [2], eCB binding to CB1R [3] and lower glutamate release [4]. g 15 min bath application of 100 µm DHPG resulted in prominent DHPG-LTD in both Pyr-WT (53.25 ± 0.8 %; n = 11 slices/ 4 mice; p < 0.001 relative to baseline; one-way Anova) and Pyr-KO (45.1 ± 0.80%; n = 11 slices/ 4 mice; p < 0.001 relative to baseline; one-way Anova). h Average PPR after DHPG application. During DHPG-LTD (30–90 min post DHPG), PPR in Pyr-WT: 1.89 ± 0.16 (n = 11 slices, 4 mice) vs. PPR in Pyr-KO: 2.47 ± 0.35 (n = 11 slices, 4 mice); p = 0.621. 2-Way RM Anova. Insets: fEPSP trace examples from Pyr-WT and Pyr-KO slices before (gray line) and after induction (black line) of WIN-LTD (d) or DHPG-LTD (g); each trace is average of 30 individual traces taken at baseline or at 75–90 min post-induction.