Journal: bioRxiv

Article Title: Enhanced CB1 receptor function in GABAergic neurons mediates hyperexcitability and impaired sensory-driven synchrony of cortical circuits in Fragile X Syndrome model mice

doi: 10.1101/2025.01.02.630932

Figure Lengend Snippet: A) Up states were recorded extracellularly in layer 4 of somatosensory cortex. B) Example traces collected in vehicle and the CBR1 antagonist, Rimonabant (5 µM). C) Bar-graph of Up state duration showing a prolonged phenotype in vehicle and no detectable change in the Fmr1 KO with Rimonabant application (N=20-42 slices, 5-9 mice for each condition). D) Example traces collected in vehicle and the diacylglycerol lipase inhibitor, DO34 (10 µM). E) Bar-graph of Up state duration showing a prolonged phenotype in vehicle and a partial rescue in the Fmr1 KO with DO34 application (N=21-29 slices and 5-7 mice for each condition). Scale bars for B and D = 50 µV and 1 s. 2-way ANOVA with Sidak’s multiple comparisons test. F,G) Enhanced CB1R-mediated suppression of inhibitory synaptic transmission in Fmr1 KO cortex. F) Examples mIPSCs from a L2/3 pyramidal neuron before (baseline) and after Rimonabant (5 µM) application. Scale bars = 5 pA and 50 ms. B) Average mIPSC frequency (normalized to pre-rimonabant baseline) in WT and Fmr1 KO neurons. (N=14-20 cells from 7-15 mice per genotype). Repeated measures 2 way ANOVA; Rimonabant X genotype interaction (F (3, 96) = 3.366; p<0.05); Sidak’s posthoc tests; *, p<0.05 **, p<0.01 ****, p<0.001.

Article Snippet: Rimonabant (SR141716A, 5 µM, Tocris, Cat 0923.

Techniques: Transmission Assay

Journal: bioRxiv

Article Title: Enhanced CB1 receptor function in GABAergic neurons mediates hyperexcitability and impaired sensory-driven synchrony of cortical circuits in Fragile X Syndrome model mice

doi: 10.1101/2025.01.02.630932

Figure Lengend Snippet: A) Diagram of EEG electrode array laid over scalp of mice. B) Chirp stimuli were 2 seconds of broadband noise sinusoidally modulated starting at 1 Hz and ramping up to 100 Hz. C) Cnr1 genetic reduction experiment. Top : ITPC average plots of induced activity in the right medial region (red box in A) in the 3 genotypes examined (WT, Fmr1 KO, and Fmr1 KO with heterozygous deletion of Cnr1 in GABAergic neurons; VGAT Cre / Cnr1 fl/+ / Fmr1 KO). Bottom : Average difference plots (ΔITPC) based on plots above indicate decreased ITPC at higher frequencies in Fmr1 KO mice (left) and the rescue of this phenotype with a genetic reduction of Cnr1 in GABAergic neurons . N=17,17,17 mice. D) Rimonabant treatment experiment. Left : ITPC average plots of induced activity for pre- and post-rimonabant treatment obtained from the left medial region (blue box in A) in Fmr1 KO mice (N=14). Right : Difference plot indicating increases in ITPC with rimonabant treatment. Solid lines border statistically different areas of the plot. Scales in C apply to D.

Article Snippet: Rimonabant (SR141716A, 5 µM, Tocris, Cat 0923.

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Enhanced CB1 receptor function in GABAergic neurons mediates hyperexcitability and impaired sensory-driven synchrony of cortical circuits in Fragile X Syndrome model mice

doi: 10.1101/2025.01.02.630932

Figure Lengend Snippet: A) Diagram of EEG electrode array. Data shown are from the left frontal region electrodes (red box). B) A 40 Hz pulse train was used to induce an ASSR. C) Genetic reduction experiment. Top : ITPC average plots of induced activity in the 3 genotypes examined. Bottom : Average difference plots indicate decreased ITPC during the ASSR in Fmr1 KO mice (left) and the rescue of this phenotype in Fmr1 KO with genetic reduction of Cnr1 in GABAergic neurons (VGAT Cre / Cnr1 fl/+ / Fmr1 KO). N=17,17,17 mice. D) Rimonabant treatment experiment. Left : ITPC average plots of induced activity for pre- and post-rimonabant treatment obtained from the left medial region in Fmr1 KO mice (N=14). Right : Difference plot indicating increases in ITPC with rimonabant treatment. Solid lines border statistically different areas of the plot. Scales in C apply to D.

Article Snippet: Rimonabant (SR141716A, 5 µM, Tocris, Cat 0923.

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Enhanced CB1 receptor function in GABAergic neurons mediates hyperexcitability and impaired sensory-driven synchrony of cortical circuits in Fragile X Syndrome model mice

doi: 10.1101/2025.01.02.630932

Figure Lengend Snippet: A) Diagram of EEG electrode array. Data shown are from the left temporal region electrodes (red box). B) An 80 Hz pulse train was used to induce an ASSR (auditory steady-state response). C) Top : ITPC average plots of induced activity in the 3 genotypes examined. Bottom : Average difference plots indicate decreased ITPC during the ASSR in Fmr1 KO mice (left) and the rescue of this phenotype in Fmr1 KO mice with genetic reduction of Cnr1 in GABAergic neurons (VGAT Cre / Cnr1 fl/+ / Fmr1 KO). N=17,17,17 mice. D) Left : ITPC average plots of induced activity for pre- and post-rimonabant treatment in Fmr1 KO mice (N=14). Right : Difference plot indicating an increase in ITPC with rimonabant treatment. Solid lines border statistically different areas of the plot. Scales in C apply to D.

Article Snippet: Rimonabant (SR141716A, 5 µM, Tocris, Cat 0923.

Techniques: Activity Assay

Journal: bioRxiv

Article Title: Enhanced CB1 receptor function in GABAergic neurons mediates hyperexcitability and impaired sensory-driven synchrony of cortical circuits in Fragile X Syndrome model mice

doi: 10.1101/2025.01.02.630932

Figure Lengend Snippet: Spectral density power (normalized to wild-type (WT) mice (A) or to pre-treatment Fmr1 KO mice (B)) is plotted for 6 standard frequency bands and obtained from the right temporal electrode group (as indicated by red box in lower right). A) An increase in power is observed in the low and high gamma frequency bands in the Fmr1 KO mice ( black ) and as compared to WT littermates. Fmr1 KO mice with genetic reduction of Cnr1 in GABAergic neurons (VGAT Cre/ Cnr1 fl/+ /Fmr1 KO) (gray) have increased power across all frequency bands, as compared to WT, and is not different from Fmr1 KO, with the exception of high gamma. N=17, 17, 17 mice. B) 7 day rimonabant treatment of Fmr1 KO mice decreases power in all frequency bands except High Gamma. *, p<0.05 **, p<0.01 ****, p<0.001

Article Snippet: Rimonabant (SR141716A, 5 µM, Tocris, Cat 0923.

Techniques:

Journal: Addiction Biology

Article Title: Interrelated involvement of the endocannabinoid/endovanilloid (TRPV1) systems and epigenetic processes in anxiety‐ and working memory impairment‐related behavioural effects of nicotine as a stressor

doi: 10.1111/adb.13421

Figure Lengend Snippet: Mitigating effects of the HDAC inhibitors, CB1 agonist, or TRPV1 antagonist against anxiety‐like behaviours. The parameter values of the EPM test (percentages of entries into open arms and time spent on open arms) at the 2‐h time point after the last NC (0.8 mg/kg, sc) and/or IM (10 min) treatment are shown as means with standard deviation (SD) bars ( n = 10) for each HDAC inhibitor (SB or VA), CB1 agonist (AC), or TRPV1 antagonist (CZ) cotreatment group (with each ip dose [mg/kg]), and statistical significance in posthoc tests is denoted using the symbols as defined below. The detailed data and statistical results have been included in Table . (A) SB (100 mg/kg, ip) cotreatment groups (SB groups); (B) VA (300 mg/kg, ip) cotreatment groups (VA groups); (C) AC (0.2 mg/kg, ip) cotreatment groups (AC groups); (D) CZ (1 mg/kg, ip) cotreatment groups (CZ groups). ** P < 0.01: significant attenuation as compared with the control group; ++ P < 0.01: significant increase as compared with the NC, IM, or NC‐IM group without any cotreatments; $$ P < 0.01: significant attenuation as compared with the IM group without any cotreatments.

Article Snippet: In the NC treatment groups, repeated subcutaneous (sc) doses of NC that caused both anxiety‐ and working memory impairment‐like behaviours effectively in mice were selected: a single sc dose of 0.8 mg/kg was administered daily for 4 days., NC (Nacalai Tesque, Inc., Kyoto, Japan) was supplied in free‐base form at 95% purity, and was freshly dissolved in saline to a volume of 5 mL/kg immediately before each administration., , For the stressor treatment groups, repeated immobilization (IM) stress treatments in which anxiogenic‐ and working memory impairment‐like effects similar to those of the NC treatments were selected: 10 min of IM, which was induced by placing the mouse in a narrow space (diameter about 12 cm) in a vinyl bag with some breathing holes, was performed once a day for 4 days., Furthermore, to investigate the interactions between NC and IM, the behavioural alterations were examined in the NC plus IM group (NC‐IM group) which received the aforementioned sc dose of NC 10 min before the IM treatment once a day for 4 days., , With respect to the HDAC inhibitors sodium butyrate (SB) and valproic acid (VA), the cannabinoid type 1 (CB1) agonist ACPA (arachidonylcyclopropylamide [AC]), the CB1 antagonist SR 141716A ( N ‐(piperidin‐1‐yl)‐5‐(4‐chlorophenyl)‐1‐(2,4‐dichlorophenyl)‐4‐methyl‐1H‐pyrazole‐3‐carboxamide hydrochloride [SR]), the TRPV1 agonist olvanil (OL), and the TRPV1 antagonist capsazepine (CZ), which were all purchased from Tocris Cookson Inc. (Ellisville, Missouri, USA), the data were collected and shown for the following intraperitoneal (ip) doses: 50, 100, and 200 mg/kg for SB; 200, 300, and 400 mg/kg for VA; 0.05, 0.2, and 1 mg/kg for AC; 0.5, 1, and 2 mg/kg for SR; 0.1, 1, and 2.5 mg/kg for OL; and 0.1, 1, and 5 mg/kg for CZ, based on previous data and preliminary experiments., , , , , , , , , As ligands related to the ECB system, ligands for the CB1 receptors, the representative CB receptors expressed in the central nervous system, were selected., , , , , The doses were selected from those that induced no toxic behavioural alterations (e.g., continuous suppression of locomotor activity) by themselves at the prescribed time point even when combined administration was repeated.

Techniques: Standard Deviation, Control

Journal: Addiction Biology

Article Title: Interrelated involvement of the endocannabinoid/endovanilloid (TRPV1) systems and epigenetic processes in anxiety‐ and working memory impairment‐related behavioural effects of nicotine as a stressor

doi: 10.1111/adb.13421

Figure Lengend Snippet: Mitigating effects of the HDAC inhibitors, TRPV1 agonist, or CB1 antagonist against working memory impairment‐like behaviours. The parameter values of the Y‐maze test (SAP rates) at the 2‐h time point after the last NC (0.8 mg/kg, sc) and/or IM (10 min) treatment are shown as means with SD bars ( n = 10) for each HDAC inhibitor (SB or VA), TRPV1 agonist (OL), or CB1 antagonist (SR) cotreatment group (with each ip dose [mg/kg]), and statistical significance in post hoc tests is denoted using the symbols as defined below. The detailed data and statistical results have been included in Table . (A) SB (100 mg/kg, ip) cotreatment groups (SB groups); (B) VA (300 mg/kg, ip) cotreatment groups (VA groups); (C) OL (1 mg/kg, ip) cotreatment groups (OL groups); (D) SR (1 mg/kg, ip) cotreatment groups (SR groups). ** P < 0.01: significant attenuation as compared with the control group; ++ P < 0.01: significant increase as compared with the NC, IM, or NC‐IM group without any cotreatments; @@ P < 0.01: significant attenuation as compared with the NC group without any cotreatments; $$ P < 0.01: significant attenuation as compared with the IM group without any cotreatments.

Article Snippet: In the NC treatment groups, repeated subcutaneous (sc) doses of NC that caused both anxiety‐ and working memory impairment‐like behaviours effectively in mice were selected: a single sc dose of 0.8 mg/kg was administered daily for 4 days., NC (Nacalai Tesque, Inc., Kyoto, Japan) was supplied in free‐base form at 95% purity, and was freshly dissolved in saline to a volume of 5 mL/kg immediately before each administration., , For the stressor treatment groups, repeated immobilization (IM) stress treatments in which anxiogenic‐ and working memory impairment‐like effects similar to those of the NC treatments were selected: 10 min of IM, which was induced by placing the mouse in a narrow space (diameter about 12 cm) in a vinyl bag with some breathing holes, was performed once a day for 4 days., Furthermore, to investigate the interactions between NC and IM, the behavioural alterations were examined in the NC plus IM group (NC‐IM group) which received the aforementioned sc dose of NC 10 min before the IM treatment once a day for 4 days., , With respect to the HDAC inhibitors sodium butyrate (SB) and valproic acid (VA), the cannabinoid type 1 (CB1) agonist ACPA (arachidonylcyclopropylamide [AC]), the CB1 antagonist SR 141716A ( N ‐(piperidin‐1‐yl)‐5‐(4‐chlorophenyl)‐1‐(2,4‐dichlorophenyl)‐4‐methyl‐1H‐pyrazole‐3‐carboxamide hydrochloride [SR]), the TRPV1 agonist olvanil (OL), and the TRPV1 antagonist capsazepine (CZ), which were all purchased from Tocris Cookson Inc. (Ellisville, Missouri, USA), the data were collected and shown for the following intraperitoneal (ip) doses: 50, 100, and 200 mg/kg for SB; 200, 300, and 400 mg/kg for VA; 0.05, 0.2, and 1 mg/kg for AC; 0.5, 1, and 2 mg/kg for SR; 0.1, 1, and 2.5 mg/kg for OL; and 0.1, 1, and 5 mg/kg for CZ, based on previous data and preliminary experiments., , , , , , , , , As ligands related to the ECB system, ligands for the CB1 receptors, the representative CB receptors expressed in the central nervous system, were selected., , , , , The doses were selected from those that induced no toxic behavioural alterations (e.g., continuous suppression of locomotor activity) by themselves at the prescribed time point even when combined administration was repeated.

Techniques: Control

Journal: Addiction Biology

Article Title: Interrelated involvement of the endocannabinoid/endovanilloid (TRPV1) systems and epigenetic processes in anxiety‐ and working memory impairment‐related behavioural effects of nicotine as a stressor

doi: 10.1111/adb.13421

Figure Lengend Snippet: Interacting effects between the CB1 antagonist and mitigating (anxiolytic‐like) drug (i.e., HDAC inhibitor, CB1 agonist, or TRPV1 antagonist) against anxiety‐like behavioural alterations caused by NC and/or IM. The parameter values of the EPM test at the 2‐h time point after the last NC (0.8 mg/kg, sc) or IM (10 min) treatment are shown as means with SD bars ( n = 10) for each mitigating drug plus CB1 antagonist group (with each ip dose [mg/kg]), and statistical significance in post hoc tests is denoted using the symbols as defined below. The detailed data and statistical results have been included in Table . (A) SB (100 mg/kg, ip) plus SR (1 mg/kg, ip) groups (SB + SR groups); (B) VA (300 mg/kg, ip) plus SR (1 mg/kg, ip) groups (VA + SR groups); (C) AC (0.2 mg/kg, ip) plus SR (1 mg/kg, ip) groups (AC + SR groups); (D) CZ (1 mg/kg, ip) plus SR (1 mg/kg, ip) groups (CZ + SR groups). ** P < 0.01: significant attenuation as compared with the control group; ++ P < 0.01: significant increase as compared with the NC, IM, or NC‐IM group without any cotreatments; $$ P < 0.01: significant attenuation as compared with the IM group without any cotreatments; ## P < 0.01: significant attenuation as compared with the NC, IM, or NC‐IM group cotreated with the efficacious HDAC inhibitor, CB1 agonist, or TRPV1 antagonist.

Article Snippet: In the NC treatment groups, repeated subcutaneous (sc) doses of NC that caused both anxiety‐ and working memory impairment‐like behaviours effectively in mice were selected: a single sc dose of 0.8 mg/kg was administered daily for 4 days., NC (Nacalai Tesque, Inc., Kyoto, Japan) was supplied in free‐base form at 95% purity, and was freshly dissolved in saline to a volume of 5 mL/kg immediately before each administration., , For the stressor treatment groups, repeated immobilization (IM) stress treatments in which anxiogenic‐ and working memory impairment‐like effects similar to those of the NC treatments were selected: 10 min of IM, which was induced by placing the mouse in a narrow space (diameter about 12 cm) in a vinyl bag with some breathing holes, was performed once a day for 4 days., Furthermore, to investigate the interactions between NC and IM, the behavioural alterations were examined in the NC plus IM group (NC‐IM group) which received the aforementioned sc dose of NC 10 min before the IM treatment once a day for 4 days., , With respect to the HDAC inhibitors sodium butyrate (SB) and valproic acid (VA), the cannabinoid type 1 (CB1) agonist ACPA (arachidonylcyclopropylamide [AC]), the CB1 antagonist SR 141716A ( N ‐(piperidin‐1‐yl)‐5‐(4‐chlorophenyl)‐1‐(2,4‐dichlorophenyl)‐4‐methyl‐1H‐pyrazole‐3‐carboxamide hydrochloride [SR]), the TRPV1 agonist olvanil (OL), and the TRPV1 antagonist capsazepine (CZ), which were all purchased from Tocris Cookson Inc. (Ellisville, Missouri, USA), the data were collected and shown for the following intraperitoneal (ip) doses: 50, 100, and 200 mg/kg for SB; 200, 300, and 400 mg/kg for VA; 0.05, 0.2, and 1 mg/kg for AC; 0.5, 1, and 2 mg/kg for SR; 0.1, 1, and 2.5 mg/kg for OL; and 0.1, 1, and 5 mg/kg for CZ, based on previous data and preliminary experiments., , , , , , , , , As ligands related to the ECB system, ligands for the CB1 receptors, the representative CB receptors expressed in the central nervous system, were selected., , , , , The doses were selected from those that induced no toxic behavioural alterations (e.g., continuous suppression of locomotor activity) by themselves at the prescribed time point even when combined administration was repeated.

Techniques: Control

Journal: Addiction Biology

Article Title: Interrelated involvement of the endocannabinoid/endovanilloid (TRPV1) systems and epigenetic processes in anxiety‐ and working memory impairment‐related behavioural effects of nicotine as a stressor

doi: 10.1111/adb.13421

Figure Lengend Snippet: Counteraction caused by the TRPV1 antagonist against attenuating effects of the CB1 antagonist on HDAC inhibitor‐ or CB1 agonist‐induced anxiolytic‐like behavioural alterations in the NC and/or IM treatment groups. The parameter values of the EPM test at the 2‐h time point after the last NC (0.8 mg/kg, sc) and/or IM (10 min) treatment are shown as means with SD bars ( n = 10) for each anxiolytic‐like drug (HDAC inhibitor or CB1 agonist) plus CB1 antagonist cotreatment group with or without additional TRPV1 antagonist (with each ip dose [mg/kg]), and statistical significance in post hoc tests is denoted using the symbols as defined below. The detailed data and statistical results have been included in Table . (A) SB (100 mg/kg, ip) plus SR (1 mg/kg, ip) cotreatment groups with (or without) additional CZ (1 mg/kg, ip) (SB + SR + CZ groups); (B) VA (300 mg/kg, ip) plus SR (1 mg/kg, ip) cotreatment groups with (or without) additional CZ (1 mg/kg, ip) (VA + SR + CZ groups); (C) AC (0.2 mg/kg, ip) plus SR (1 mg/kg, ip) cotreatment groups with (or without) additional CZ (1 mg/kg, ip) (AC + SR + CZ groups). ** P < 0.01: significant attenuation as compared with the control group; ++ P < 0.01: significant increase as compared with the NC, IM, or NC‐IM group without any cotreatments; $$ P < 0.01: significant attenuation as compared with the IM group without any cotreatments; ## P < 0.01: significant attenuation as compared with the NC, IM, or NC‐IM group cotreated with the efficacious (anxiolytic‐like) HDAC inhibitor or CB1 agonist; && P < 0.01: significant increase as compared with the NC, IM, or NC‐IM group cotreated with the mitigating drug (HDAC inhibitor or CB1 agonist) plus SR.

Article Snippet: In the NC treatment groups, repeated subcutaneous (sc) doses of NC that caused both anxiety‐ and working memory impairment‐like behaviours effectively in mice were selected: a single sc dose of 0.8 mg/kg was administered daily for 4 days., NC (Nacalai Tesque, Inc., Kyoto, Japan) was supplied in free‐base form at 95% purity, and was freshly dissolved in saline to a volume of 5 mL/kg immediately before each administration., , For the stressor treatment groups, repeated immobilization (IM) stress treatments in which anxiogenic‐ and working memory impairment‐like effects similar to those of the NC treatments were selected: 10 min of IM, which was induced by placing the mouse in a narrow space (diameter about 12 cm) in a vinyl bag with some breathing holes, was performed once a day for 4 days., Furthermore, to investigate the interactions between NC and IM, the behavioural alterations were examined in the NC plus IM group (NC‐IM group) which received the aforementioned sc dose of NC 10 min before the IM treatment once a day for 4 days., , With respect to the HDAC inhibitors sodium butyrate (SB) and valproic acid (VA), the cannabinoid type 1 (CB1) agonist ACPA (arachidonylcyclopropylamide [AC]), the CB1 antagonist SR 141716A ( N ‐(piperidin‐1‐yl)‐5‐(4‐chlorophenyl)‐1‐(2,4‐dichlorophenyl)‐4‐methyl‐1H‐pyrazole‐3‐carboxamide hydrochloride [SR]), the TRPV1 agonist olvanil (OL), and the TRPV1 antagonist capsazepine (CZ), which were all purchased from Tocris Cookson Inc. (Ellisville, Missouri, USA), the data were collected and shown for the following intraperitoneal (ip) doses: 50, 100, and 200 mg/kg for SB; 200, 300, and 400 mg/kg for VA; 0.05, 0.2, and 1 mg/kg for AC; 0.5, 1, and 2 mg/kg for SR; 0.1, 1, and 2.5 mg/kg for OL; and 0.1, 1, and 5 mg/kg for CZ, based on previous data and preliminary experiments., , , , , , , , , As ligands related to the ECB system, ligands for the CB1 receptors, the representative CB receptors expressed in the central nervous system, were selected., , , , , The doses were selected from those that induced no toxic behavioural alterations (e.g., continuous suppression of locomotor activity) by themselves at the prescribed time point even when combined administration was repeated.

Techniques: Control

Journal: Addiction Biology

Article Title: Interrelated involvement of the endocannabinoid/endovanilloid (TRPV1) systems and epigenetic processes in anxiety‐ and working memory impairment‐related behavioural effects of nicotine as a stressor

doi: 10.1111/adb.13421

Figure Lengend Snippet: Interacting effects between the TRPV1 antagonist and mitigating (working memory improving‐like) drug (i.e., HDAC inhibitor, TRPV1 agonist, or CB1 antagonist) against working memory impairment‐like behavioural alterations caused by NC and/or IM. The parameter values of the Y‐maze test at the 2‐h time point after the last NC (0.8 mg/kg, sc) or IM (10 min) treatment are shown as means with SD bars ( n = 10) for each mitigating drug plus TRPV1 antagonist group (with each ip dose [mg/kg]), and statistical significance in post hoc tests is denoted using the symbols as defined below. The detailed data and statistical results have been included in Table . (A) SB (100 mg/kg, ip) plus CZ (1 mg/kg, ip) groups (SB + CZ groups); (B) VA (300 mg/kg, ip) plus CZ (1 mg/kg, ip) groups (VA + CZ groups); (C) OL (1 mg/kg, ip) plus CZ (1 mg/kg, ip) groups (OL + CZ groups); (D) SR (1 mg/kg, ip) plus CZ (1 mg/kg, ip) groups (SR + CZ groups). ** P < 0.01: significant attenuation as compared with the control group; ++ P < 0.01: significant increase as compared with the NC, IM, or NC‐IM group without any cotreatments; @@ P < 0.01: significant attenuation as compared with the NC group without any cotreatments; $$ P < 0.01: significant attenuation as compared with the IM group without any cotreatments; ## P < 0.01: significant attenuation as compared with the NC, IM, or NC‐IM group cotreated with the efficacious HDAC inhibitor, TRPV1 agonist, or CB1 antagonist.

Article Snippet: In the NC treatment groups, repeated subcutaneous (sc) doses of NC that caused both anxiety‐ and working memory impairment‐like behaviours effectively in mice were selected: a single sc dose of 0.8 mg/kg was administered daily for 4 days., NC (Nacalai Tesque, Inc., Kyoto, Japan) was supplied in free‐base form at 95% purity, and was freshly dissolved in saline to a volume of 5 mL/kg immediately before each administration., , For the stressor treatment groups, repeated immobilization (IM) stress treatments in which anxiogenic‐ and working memory impairment‐like effects similar to those of the NC treatments were selected: 10 min of IM, which was induced by placing the mouse in a narrow space (diameter about 12 cm) in a vinyl bag with some breathing holes, was performed once a day for 4 days., Furthermore, to investigate the interactions between NC and IM, the behavioural alterations were examined in the NC plus IM group (NC‐IM group) which received the aforementioned sc dose of NC 10 min before the IM treatment once a day for 4 days., , With respect to the HDAC inhibitors sodium butyrate (SB) and valproic acid (VA), the cannabinoid type 1 (CB1) agonist ACPA (arachidonylcyclopropylamide [AC]), the CB1 antagonist SR 141716A ( N ‐(piperidin‐1‐yl)‐5‐(4‐chlorophenyl)‐1‐(2,4‐dichlorophenyl)‐4‐methyl‐1H‐pyrazole‐3‐carboxamide hydrochloride [SR]), the TRPV1 agonist olvanil (OL), and the TRPV1 antagonist capsazepine (CZ), which were all purchased from Tocris Cookson Inc. (Ellisville, Missouri, USA), the data were collected and shown for the following intraperitoneal (ip) doses: 50, 100, and 200 mg/kg for SB; 200, 300, and 400 mg/kg for VA; 0.05, 0.2, and 1 mg/kg for AC; 0.5, 1, and 2 mg/kg for SR; 0.1, 1, and 2.5 mg/kg for OL; and 0.1, 1, and 5 mg/kg for CZ, based on previous data and preliminary experiments., , , , , , , , , As ligands related to the ECB system, ligands for the CB1 receptors, the representative CB receptors expressed in the central nervous system, were selected., , , , , The doses were selected from those that induced no toxic behavioural alterations (e.g., continuous suppression of locomotor activity) by themselves at the prescribed time point even when combined administration was repeated.

Techniques: Control

Journal: Addiction Biology

Article Title: Interrelated involvement of the endocannabinoid/endovanilloid (TRPV1) systems and epigenetic processes in anxiety‐ and working memory impairment‐related behavioural effects of nicotine as a stressor

doi: 10.1111/adb.13421

Figure Lengend Snippet: Counteraction caused by the CB1 antagonist against attenuating effects of the TRPV1 antagonist on HDAC inhibitor‐ or TRPV1 agonist‐induced working memory improving‐like behavioural alterations in the NC and/or IM treatment groups. The parameter values of the Y‐maze test at the 2‐h time point after the last NC (0.8 mg/kg, sc) and/or IM (10 min) treatment are shown as means with SD bars ( n = 10) for each working memory improving‐like drug (HDAC inhibitor or TRPV1 agonist) plus TRPV1 antagonist cotreatment group with or without additional CB1 antagonist (with each ip dose [mg/kg]), and statistical significance in post hoc tests is denoted using the symbols as defined below. The detailed data and statistical results have been included in Table . (A) SB (100 mg/kg, ip) plus CZ (1 mg/kg, ip) cotreatment groups with (or without) additional SR (1 mg/kg, ip) (SB + CZ + SR groups); (B) VA (300 mg/kg, ip) plus CZ (1 mg/kg, ip) cotreatment groups with (or without) additional SR (1 mg/kg, ip) (VA + CZ + SR groups); (C) OL (1 mg/kg, ip) plus CZ (1 mg/kg, ip) cotreatment groups with (or without) additional SR (1 mg/kg, ip) (OL + CZ + SR groups). ** P < 0.01: significant attenuation as compared with the control group; ++ P < 0.01: significant increase as compared with the NC, IM, or NC‐IM group without any cotreatments; @@ P < 0.01: significant attenuation as compared with the NC group without any cotreatments; $$ P < 0.01: significant attenuation as compared with the IM group without any cotreatments; ## P < 0.01: significant attenuation as compared with the NC, IM, or NC‐IM group cotreated with the efficacious (working memory improving‐like) HDAC inhibitor or TRPV1 agonist; && P < 0.01: significant increase as compared with the NC, IM, or NC‐IM group cotreated with the mitigating drug (HDAC inhibitor or TRPV1 agonist) plus CZ.

Article Snippet: In the NC treatment groups, repeated subcutaneous (sc) doses of NC that caused both anxiety‐ and working memory impairment‐like behaviours effectively in mice were selected: a single sc dose of 0.8 mg/kg was administered daily for 4 days., NC (Nacalai Tesque, Inc., Kyoto, Japan) was supplied in free‐base form at 95% purity, and was freshly dissolved in saline to a volume of 5 mL/kg immediately before each administration., , For the stressor treatment groups, repeated immobilization (IM) stress treatments in which anxiogenic‐ and working memory impairment‐like effects similar to those of the NC treatments were selected: 10 min of IM, which was induced by placing the mouse in a narrow space (diameter about 12 cm) in a vinyl bag with some breathing holes, was performed once a day for 4 days., Furthermore, to investigate the interactions between NC and IM, the behavioural alterations were examined in the NC plus IM group (NC‐IM group) which received the aforementioned sc dose of NC 10 min before the IM treatment once a day for 4 days., , With respect to the HDAC inhibitors sodium butyrate (SB) and valproic acid (VA), the cannabinoid type 1 (CB1) agonist ACPA (arachidonylcyclopropylamide [AC]), the CB1 antagonist SR 141716A ( N ‐(piperidin‐1‐yl)‐5‐(4‐chlorophenyl)‐1‐(2,4‐dichlorophenyl)‐4‐methyl‐1H‐pyrazole‐3‐carboxamide hydrochloride [SR]), the TRPV1 agonist olvanil (OL), and the TRPV1 antagonist capsazepine (CZ), which were all purchased from Tocris Cookson Inc. (Ellisville, Missouri, USA), the data were collected and shown for the following intraperitoneal (ip) doses: 50, 100, and 200 mg/kg for SB; 200, 300, and 400 mg/kg for VA; 0.05, 0.2, and 1 mg/kg for AC; 0.5, 1, and 2 mg/kg for SR; 0.1, 1, and 2.5 mg/kg for OL; and 0.1, 1, and 5 mg/kg for CZ, based on previous data and preliminary experiments., , , , , , , , , As ligands related to the ECB system, ligands for the CB1 receptors, the representative CB receptors expressed in the central nervous system, were selected., , , , , The doses were selected from those that induced no toxic behavioural alterations (e.g., continuous suppression of locomotor activity) by themselves at the prescribed time point even when combined administration was repeated.

Techniques: Control