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. 2022 Mar 3;42(9):1650–1665. doi: 10.1177/0271678X221080324

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© The Author(s) 2022

This article is distributed under the terms of the Creative Commons Attribution 4.0 License (https://creativecommons.org/licenses/by/4.0/) which permits any use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).

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Figure 4. — Granule cells from mice decrease spontaneous and mini EPSCs and firing frequency after activation of HCAR1. (a) Example traces of sEPSCs measured during baseline condition (black) and following application of 40 µM 3Cl-HBA (red). (b) Cumulative distributions of sEPSCs frequency in WT and HCAR1 KO were analyzed by using Kolmogorov-Smirnov test. WT: P < 0.0001 versus HCAR1 activation; HCAR1 KO: P = P = 0.18 versus HCAR1 activation (c) Summary of recorded cells showing a decreased frequency of sEPSC by 36% induced by 3Cl-HBA compared to baseline (one sample t, n = 5, mean = 63.97 ± 16.79, P = 0.0087). HCAR1 KO recorded cells showed no significant decrease in sEPSC frequency after 3Cl-HBA (one sample t, n = 9, mean = 103 ± 12.58, P = 0.5) (d) sEPSC amplitude showed no statistically significant changes between baseline and 3Cl-HBA application in both WT and HCAR1 KO (WT: one sample t, n = 5, mean = 99.22 ± 12.72, P = 0.9; HCAR1 KO: one sample t, n = 9, mean = 100.1 ± 16.07, P = 0.98). (e) Cumulative distributions of mEPSCs frequency in WT and HCAR1 KO were analyzed by using Kolmogorov-Smirnov test. WT: P < 0,0001 versus HCAR1 activation; HCAR1 KO: P = 0.14 versus HCAR1 activation. (f) Summary of recorded cells showing a decreased frequency of mEPSC by 53% induced by 3Cl-HBA compared to baseline (one sample t, n = 5, mean = 46.5 ± 13.42, P = 0.0009). HCAR1 KO recorded cells showed no significant decrease in mEPSC frequency after 3Cl-HBA (one sample t, n = 6, mean = 103.9 ± 19.55, P = 0.64) (g) mEPSC amplitude showed no statistically significant changes between baseline and 3Cl-HBA application both in WT and HCAR1 KO (WT: one sample t, n = 5, mean = 96.34 ± 15.52, P = 0.63; HCAR1 KO: one sample t test, n = 6, mean = 108.1 ± 25.10, P = 0.47). (h, i) Effect of HCAR1 activation on neuronal firing frequency following steps of current injection in WT mice. The maximum number of action potentials evoked was significantly decreased by HCAR1 activation (paired t test, n = 7, Baseline: mean = 36 ± 12.58, 3Cl-HBA: mean = 29 ± 13.98, P = 0.019), but not the current injected to reach the maximum firing frequency (paired t test, n = 7, Baseline: mean = 184.3 ± 121.8, 3Cl-HBA: mean = 162.9 ± 127.1, P = 0.14). (j, k) In HCAR1 KO, activation of HCAR1 did not change the maximum number of AP compared to baseline (paired t test, n = 8, Baseline: mean = 36.75 ± 14.62, 3Cl-HBA: mean = 38.75 ± 16.06, P = 0.62), nor the current injected to reach the maximum firing frequency (paired t test, n = 8, Baseline: mean = 202.5 ± 41.66, 3Cl-HBA: mean = 198.8 ± 35.63, P = 0.82). Values are means±SD, *P < 0.05, **P < 0.01, ***P < 0.001 versus 3Cl-HBA application.