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Med Associates Inc fear conditioning box

RGS6 in DG NPCs is required for running-induced cognitive improvements in APP SWE mice. (A) Schematic outlining experimental design. Mice were placed in cages containing either fixed (control) or free (running) wheels for 8 weeks. Mice underwent hippocampal-based learning and memory behavioral tasks after 4 weeks (Y-maze [B]) and 8 weeks (Y-maze [C] and contextual fear <t>conditioning</t> (CFC, [D]) of wheel treatment. (B, C) Multi-way analysis of variance with Tukey’s post hoc adjustment was used to analyze the effects of and interactions between genotype, sex, time, and wheel treatment after 4 (B) or 8 weeks (C) of wheel treatment. Top: % Spontaneous alternations— Significant effects of genotype ( F 3,154 = 16.261, P = 0.000) and wheel treatment ( F 1,154 = 52.906, P = 0.000), as well as their interaction ( F 3,154 = 19.638, P = 0.000) were observed. No significant effects of sex or time were observed. Bottom: % Time spent in novel arm— Significant effects of genotype ( F 3,154 = 30.835, P = 0.000) and wheel treatment ( F 1,154 = 22.496, P = 0.000), as well as their interaction ( F 3,154 = 7.652, P = 0.000) were observed. No significant effects of sex or time were observed. (D) Multi-way ANOVA with Tukey’s post hoc adjustment was used to analyze the effects of and interactions between genotype, sex, and wheel treatment. A significant effect of genotype ( F 3,55 = 6.217, P = 0.001) and its interaction with wheel treatment ( F 3,55 = 3.898, P = 0.013) were observed. No significant effects of sex or wheel treatment were observed. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. (Fixed wheel: n = 8 RGS6 WT DG mice; n = 11 RGS6 KO DG mice; n = 10 RGS6 WT DG ; APP SWE mice; n = 12 RGS6 KO DG ; APP SWE mice. Free wheel: n = 7 RGS6 WT DG mice; n = 9 RGS6 KO DG mice; n = 10 RGS6 WT DG ; APP SWE mice; n = 15 RGS6 KO DG ; APP SWE mice). DCX: Doublecortin; DG: dentate gyrus; GFP: green fluorescent protein; IHC: immunohistochemistry; NPCs: neuronal progenitor cells; RGS6: Regulator of G protein signaling 6.

Fear Conditioning Box, supplied by Med Associates Inc, used in various techniques. Bioz Stars score: 96/100, based on 114 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Regulator of G protein signaling 6 mediates exercise-induced recovery of hippocampal neurogenesis, learning, and memory in a mouse model of Alzheimer’s disease"

Article Title: Regulator of G protein signaling 6 mediates exercise-induced recovery of hippocampal neurogenesis, learning, and memory in a mouse model of Alzheimer’s disease

Journal: Neural Regeneration Research

doi: 10.4103/NRR.NRR-D-23-01993

Figure Legend Snippet: RGS6 in DG NPCs is required for running-induced cognitive improvements in APP SWE mice. (A) Schematic outlining experimental design. Mice were placed in cages containing either fixed (control) or free (running) wheels for 8 weeks. Mice underwent hippocampal-based learning and memory behavioral tasks after 4 weeks (Y-maze [B]) and 8 weeks (Y-maze [C] and contextual fear conditioning (CFC, [D]) of wheel treatment. (B, C) Multi-way analysis of variance with Tukey’s post hoc adjustment was used to analyze the effects of and interactions between genotype, sex, time, and wheel treatment after 4 (B) or 8 weeks (C) of wheel treatment. Top: % Spontaneous alternations— Significant effects of genotype ( F 3,154 = 16.261, P = 0.000) and wheel treatment ( F 1,154 = 52.906, P = 0.000), as well as their interaction ( F 3,154 = 19.638, P = 0.000) were observed. No significant effects of sex or time were observed. Bottom: % Time spent in novel arm— Significant effects of genotype ( F 3,154 = 30.835, P = 0.000) and wheel treatment ( F 1,154 = 22.496, P = 0.000), as well as their interaction ( F 3,154 = 7.652, P = 0.000) were observed. No significant effects of sex or time were observed. (D) Multi-way ANOVA with Tukey’s post hoc adjustment was used to analyze the effects of and interactions between genotype, sex, and wheel treatment. A significant effect of genotype ( F 3,55 = 6.217, P = 0.001) and its interaction with wheel treatment ( F 3,55 = 3.898, P = 0.013) were observed. No significant effects of sex or wheel treatment were observed. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. (Fixed wheel: n = 8 RGS6 WT DG mice; n = 11 RGS6 KO DG mice; n = 10 RGS6 WT DG ; APP SWE mice; n = 12 RGS6 KO DG ; APP SWE mice. Free wheel: n = 7 RGS6 WT DG mice; n = 9 RGS6 KO DG mice; n = 10 RGS6 WT DG ; APP SWE mice; n = 15 RGS6 KO DG ; APP SWE mice). DCX: Doublecortin; DG: dentate gyrus; GFP: green fluorescent protein; IHC: immunohistochemistry; NPCs: neuronal progenitor cells; RGS6: Regulator of G protein signaling 6.

Techniques Used: Control, Immunohistochemistry

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Journal: Neural Regeneration Research

Article Title: Regulator of G protein signaling 6 mediates exercise-induced recovery of hippocampal neurogenesis, learning, and memory in a mouse model of Alzheimer’s disease

doi: 10.4103/NRR.NRR-D-23-01993

Figure Lengend Snippet: RGS6 in DG NPCs is required for running-induced cognitive improvements in APP SWE mice. (A) Schematic outlining experimental design. Mice were placed in cages containing either fixed (control) or free (running) wheels for 8 weeks. Mice underwent hippocampal-based learning and memory behavioral tasks after 4 weeks (Y-maze [B]) and 8 weeks (Y-maze [C] and contextual fear conditioning (CFC, [D]) of wheel treatment. (B, C) Multi-way analysis of variance with Tukey’s post hoc adjustment was used to analyze the effects of and interactions between genotype, sex, time, and wheel treatment after 4 (B) or 8 weeks (C) of wheel treatment. Top: % Spontaneous alternations— Significant effects of genotype ( F 3,154 = 16.261, P = 0.000) and wheel treatment ( F 1,154 = 52.906, P = 0.000), as well as their interaction ( F 3,154 = 19.638, P = 0.000) were observed. No significant effects of sex or time were observed. Bottom: % Time spent in novel arm— Significant effects of genotype ( F 3,154 = 30.835, P = 0.000) and wheel treatment ( F 1,154 = 22.496, P = 0.000), as well as their interaction ( F 3,154 = 7.652, P = 0.000) were observed. No significant effects of sex or time were observed. (D) Multi-way ANOVA with Tukey’s post hoc adjustment was used to analyze the effects of and interactions between genotype, sex, and wheel treatment. A significant effect of genotype ( F 3,55 = 6.217, P = 0.001) and its interaction with wheel treatment ( F 3,55 = 3.898, P = 0.013) were observed. No significant effects of sex or wheel treatment were observed. Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. (Fixed wheel: n = 8 RGS6 WT DG mice; n = 11 RGS6 KO DG mice; n = 10 RGS6 WT DG ; APP SWE mice; n = 12 RGS6 KO DG ; APP SWE mice. Free wheel: n = 7 RGS6 WT DG mice; n = 9 RGS6 KO DG mice; n = 10 RGS6 WT DG ; APP SWE mice; n = 15 RGS6 KO DG ; APP SWE mice). DCX: Doublecortin; DG: dentate gyrus; GFP: green fluorescent protein; IHC: immunohistochemistry; NPCs: neuronal progenitor cells; RGS6: Regulator of G protein signaling 6.

Article Snippet: For each trial, mice are placed in a fear conditioning box (Med Associates NIR VFC System, Fairfax, VT, USA) with a steel wire bottom.

Techniques: Control, Immunohistochemistry

a Schematics of AAV injections and experimental design for habituation (day 0, D0), trace fear conditioning (TFC, D1), and retention testing (Re, D2). CS conditioned stimulus, US unconditioned stimulus. b Quantification and comparison of freezing behavior during TFC and retention across experimental and control groups. BL baseline. Associative learning groups: paired (CS-US) (green, n = 8 mice); control groups: CS only (black, n = 10 mice) and US only (yellow, n = 6 mice). These group sizes apply to all subsequent panels in this figure. (CS-US vs CS only, P = 0.00005; CS-US vs US only, P = 0.0007; CS only vs US only, P = 0.75, Mann–Whitney test (two-sided)). c , d Group-averaged Ca²⁺ responses and quantification of mPFC ChCs to pre-exposure CS during habituation across groups (CS-US: P = 0.000048, paired two-sided t -test; CS only: P = 0.00195, Wilcoxon signed-rank test (two-sided); US only: P = 0.011, paired two-sided t -test). e–g Ca²⁺ responses of mPFC ChCs to CS during TFC and retention, showing learning-dependent enhancement in the paired group ( f F 1,16 = 11.83, P = 0.0034, two-way repeated-measures ANOVA; Sidak’s multiple comparisons test (two-sided): trial #5, P = 0.006; trial #6, P = 0.005. D2 Re: CS-US vs CS only, P = 0.000061; CS-US vs US only, P = 0.0057; CS only vs US only, P = 0.28; unpaired two-sided t -test. g CS-US: P = 0.0037; CS only: P = 0.45, paired two-sided t -test). h Quantification and comparison of ChC Ca²⁺ responses to US during TFC across groups ( h F 1,12 = 8.769, P = 0.012, two-way repeated-measures ANOVA; Sidak’s multiple comparisons test (two-sided): trial #4, P = 0.022; trial #5, P = 0.034; trial #6, P = 0.004. i CS-US: P = 0.84; US only: P = 0.02, paired two-sided t -test). Each line linking two open symbols represented the data from an individual mouse. Data are presented as mean ± SEM, with individual animals shown as open symbols; box plots indicate median, quartiles, and range. Statistics as indicated, *** P < 0.001, ** P < 0.01, * P < 0.05, ns is not significant. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Prefrontal chandelier cells encode stimulus salience to influence learning in male mice

doi: 10.1038/s41467-026-68959-3

Figure Lengend Snippet: a Schematics of AAV injections and experimental design for habituation (day 0, D0), trace fear conditioning (TFC, D1), and retention testing (Re, D2). CS conditioned stimulus, US unconditioned stimulus. b Quantification and comparison of freezing behavior during TFC and retention across experimental and control groups. BL baseline. Associative learning groups: paired (CS-US) (green, n = 8 mice); control groups: CS only (black, n = 10 mice) and US only (yellow, n = 6 mice). These group sizes apply to all subsequent panels in this figure. (CS-US vs CS only, P = 0.00005; CS-US vs US only, P = 0.0007; CS only vs US only, P = 0.75, Mann–Whitney test (two-sided)). c , d Group-averaged Ca²⁺ responses and quantification of mPFC ChCs to pre-exposure CS during habituation across groups (CS-US: P = 0.000048, paired two-sided t -test; CS only: P = 0.00195, Wilcoxon signed-rank test (two-sided); US only: P = 0.011, paired two-sided t -test). e–g Ca²⁺ responses of mPFC ChCs to CS during TFC and retention, showing learning-dependent enhancement in the paired group ( f F 1,16 = 11.83, P = 0.0034, two-way repeated-measures ANOVA; Sidak’s multiple comparisons test (two-sided): trial #5, P = 0.006; trial #6, P = 0.005. D2 Re: CS-US vs CS only, P = 0.000061; CS-US vs US only, P = 0.0057; CS only vs US only, P = 0.28; unpaired two-sided t -test. g CS-US: P = 0.0037; CS only: P = 0.45, paired two-sided t -test). h Quantification and comparison of ChC Ca²⁺ responses to US during TFC across groups ( h F 1,12 = 8.769, P = 0.012, two-way repeated-measures ANOVA; Sidak’s multiple comparisons test (two-sided): trial #4, P = 0.022; trial #5, P = 0.034; trial #6, P = 0.004. i CS-US: P = 0.84; US only: P = 0.02, paired two-sided t -test). Each line linking two open symbols represented the data from an individual mouse. Data are presented as mean ± SEM, with individual animals shown as open symbols; box plots indicate median, quartiles, and range. Statistics as indicated, *** P < 0.001, ** P < 0.01, * P < 0.05, ns is not significant. Source data are provided as a Source Data file.

Article Snippet: Neutral tone (8 kHz, 60 dB, 2 s) and electric foot shock (0.6 mA, 2 s) were delivered by the standard fear conditioning shock box (UGO BASILE SRL, Italy).

Techniques: Comparison, Control, MANN-WHITNEY

a , b Schematic of the experimental design and trace fear conditioning paradigm with optogenetic inhibition during conditioning. c Group-averaged Z-scored Ca²⁺ traces (mean ± SEM) of mPFC ChCs aligned to CS or US across habituation (D0), conditioning (D1), and retention (D2) in AI eJaws and mRuby3 groups (left: AI eJaws, n = 8 mice; right: AI mRuby3, n = 8 mice, group sizes apply to all panels). CS, US, and light delivery periods are indicated in the figure. d , e Quantification and comparison of ChCs Ca²⁺ responses to the CS across D0-D2 and to the US during D1 in AI eJaws and AI mRuby3 groups ( d D0 CS, F 1, 14 = 0.2399, P = 0.6319, two-way repeated-measures ANOVA; D1 CS, F 1, 14 = 3.486, P = 0.0830; Sidak’s multiple comparisons test (two-sided): trial6, P = 0.0022. D1 US, F 1, 14 = 0.9478, P = 0.3468, two-way repeated-measures ANOVA. e AI mRuby3: D0 CS, Trial1 vs Trial6, P = 0.0001; D1 CS, Trial1 vs Trial6, P = 0.0007; D1 US, Trial1 vs Trial6, P = 0.12, paired two-sided t -test. AI eJaws: D0 CS, Trial1 vs Trial6, P = 0.00088; D1 CS, Trial1 vs Trial6, P = 0.28, paired two-sided t -test; D1 US, Trial1 vs Trial6, P = 0.012, paired two-sided t -test. AI eJaws vs AI mRuby3: D1 CS trial6, P = 0.00002; D2 CS, P = 0.012; D1 US trial6, P = 0.07, unpaired two-sided t -test). f–h Same as ( c–e ), but for PVT eJaws and PVT mRuby3 groups ( g D0 CS, F 1, 16 = 0.0074, P = 0.9325, two-way repeated-measures ANOVA; D1 CS, F 1, 16 = 1.929, P = 0.1839, two-way repeated-measures ANOVA; Sidak’s multiple comparisons test (two-sided): trial6, P = 0.0087; D1 US, F 1, 16 = 0.4206, P = 0.5259, two-way repeated-measures ANOVA. h PVT mRuby3: D0 CS, Trial1 vs Trial6, P = 0.00007; D1 CS, Trial1 vs Trial6, P = 0.00005, paired two-sided t -test; D1 US, Trial1 vs Trial6, P = 0.30, Wilcoxon signed-rank test (two-sided). PVT eJaws: D0 CS, Trial1 vs Trial6, P = 0.000075; D1 CS, Trial1 vs Trial6, P = 0.53; D1 US, Trial1 vs Trial6, P = 0.033, paired two-sided t -test. PVT eJaws vs PVT mRuby3: D1 CS trial6, P = 0.00045; D2 CS, P = 0.000003, unpaired two-sided t -test; D1 US trial6, P = 0.013, Mann–Whitney test (two-sided); PVT eJaws, n = 9 mice, PVT mRuby3, n = 9 mice). i Behavioral effects of optogenetic inhibition of AI inputs during TFC. Left: Freezing behavior across D1 (conditioning) and D2 (retention. F 1, 14 = 4.296, P = 0.0571, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial 4, P = 0.0375, trial 5, P = 0.0244, trial 6, P = 0.0089) Right: Comparison of freezing responses to CS during D2 (AI eJaws vs AI mRuby3, P = 0.014, unpaired two-sided t -test). j Same as ( i ), but for PVT inhibition (D1: F 1, 16 = 1.18, P = 0.29, two-way repeated-measures ANOVA; D2, PVT eJaws vs PVT mRuby3, P = 0.006, Mann–Whitney test (two-sided); PVT mRuby3 vs AI mRuby3, P = 0.77, unpaired two-sided t -test). Statistics as indicated, *** P < 0.001, ** P < 0.01, * P < 0.05, ns is not significant. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Prefrontal chandelier cells encode stimulus salience to influence learning in male mice

doi: 10.1038/s41467-026-68959-3

Figure Lengend Snippet: a , b Schematic of the experimental design and trace fear conditioning paradigm with optogenetic inhibition during conditioning. c Group-averaged Z-scored Ca²⁺ traces (mean ± SEM) of mPFC ChCs aligned to CS or US across habituation (D0), conditioning (D1), and retention (D2) in AI eJaws and mRuby3 groups (left: AI eJaws, n = 8 mice; right: AI mRuby3, n = 8 mice, group sizes apply to all panels). CS, US, and light delivery periods are indicated in the figure. d , e Quantification and comparison of ChCs Ca²⁺ responses to the CS across D0-D2 and to the US during D1 in AI eJaws and AI mRuby3 groups ( d D0 CS, F 1, 14 = 0.2399, P = 0.6319, two-way repeated-measures ANOVA; D1 CS, F 1, 14 = 3.486, P = 0.0830; Sidak’s multiple comparisons test (two-sided): trial6, P = 0.0022. D1 US, F 1, 14 = 0.9478, P = 0.3468, two-way repeated-measures ANOVA. e AI mRuby3: D0 CS, Trial1 vs Trial6, P = 0.0001; D1 CS, Trial1 vs Trial6, P = 0.0007; D1 US, Trial1 vs Trial6, P = 0.12, paired two-sided t -test. AI eJaws: D0 CS, Trial1 vs Trial6, P = 0.00088; D1 CS, Trial1 vs Trial6, P = 0.28, paired two-sided t -test; D1 US, Trial1 vs Trial6, P = 0.012, paired two-sided t -test. AI eJaws vs AI mRuby3: D1 CS trial6, P = 0.00002; D2 CS, P = 0.012; D1 US trial6, P = 0.07, unpaired two-sided t -test). f–h Same as ( c–e ), but for PVT eJaws and PVT mRuby3 groups ( g D0 CS, F 1, 16 = 0.0074, P = 0.9325, two-way repeated-measures ANOVA; D1 CS, F 1, 16 = 1.929, P = 0.1839, two-way repeated-measures ANOVA; Sidak’s multiple comparisons test (two-sided): trial6, P = 0.0087; D1 US, F 1, 16 = 0.4206, P = 0.5259, two-way repeated-measures ANOVA. h PVT mRuby3: D0 CS, Trial1 vs Trial6, P = 0.00007; D1 CS, Trial1 vs Trial6, P = 0.00005, paired two-sided t -test; D1 US, Trial1 vs Trial6, P = 0.30, Wilcoxon signed-rank test (two-sided). PVT eJaws: D0 CS, Trial1 vs Trial6, P = 0.000075; D1 CS, Trial1 vs Trial6, P = 0.53; D1 US, Trial1 vs Trial6, P = 0.033, paired two-sided t -test. PVT eJaws vs PVT mRuby3: D1 CS trial6, P = 0.00045; D2 CS, P = 0.000003, unpaired two-sided t -test; D1 US trial6, P = 0.013, Mann–Whitney test (two-sided); PVT eJaws, n = 9 mice, PVT mRuby3, n = 9 mice). i Behavioral effects of optogenetic inhibition of AI inputs during TFC. Left: Freezing behavior across D1 (conditioning) and D2 (retention. F 1, 14 = 4.296, P = 0.0571, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial 4, P = 0.0375, trial 5, P = 0.0244, trial 6, P = 0.0089) Right: Comparison of freezing responses to CS during D2 (AI eJaws vs AI mRuby3, P = 0.014, unpaired two-sided t -test). j Same as ( i ), but for PVT inhibition (D1: F 1, 16 = 1.18, P = 0.29, two-way repeated-measures ANOVA; D2, PVT eJaws vs PVT mRuby3, P = 0.006, Mann–Whitney test (two-sided); PVT mRuby3 vs AI mRuby3, P = 0.77, unpaired two-sided t -test). Statistics as indicated, *** P < 0.001, ** P < 0.01, * P < 0.05, ns is not significant. Source data are provided as a Source Data file.

Article Snippet: Neutral tone (8 kHz, 60 dB, 2 s) and electric foot shock (0.6 mA, 2 s) were delivered by the standard fear conditioning shock box (UGO BASILE SRL, Italy).

Techniques: Inhibition, Comparison, MANN-WHITNEY

a Viral injection, fiber implantation, and representative NpHR expression in mPFC ChCs. b , c Behavioral effects of ChC inhibition during conditioning, applied as indicated (NpHR group (orange) vs EYFP group (black): D1 TFC, F 1,17 = 21.57, P = 0.0002, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial #3, P = 0.0087, trial #4, P = 0.0052, trial #5, P < 0.0001, trial #6, P = 0.0348. D2 Re, NpHR vs EYFP, P = 0.0047, unpaired two-sided t -test. NpHR, n = 11 mice; EYFP, n = 8 mice). c D1 TFC, F 1,16 = 11.69, P = 0.0035, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial #3, P = 0.0495, trial #6, P = 0.0153. D2 Re, NpHR vs EYFP, P = 0.0038, unpaired two-sided t -test. NpHR, n = 9 mice; EYFP, n = 9 mice). d , e Schematics of AAV injections and experimental design for chemogenetic manipulation of ChC activity, with representative images of GCaMP7s co-expressed with hM3Dq ( n = 7 mice) or hM4Di ( n = 9 mice). f , g Corresponding average traces and quantification of ChC Ca²⁺ responses to tone stimuli at baseline and after CNO ( f hM3Dq, P = 0.0029, paired two-sided t -test, n = 7 mice. g hM4Di, P = 0.0075, paired two-sided t -test, n = 9 mice). h Schematics and representative hM3Dq expression in mPFC ChCs. i , j Behavioral effects of chemogenetic activation or inhibition of ChCs during conditioning ( i CNO(hM3Dq) vs Saline, D1 TFC, F 1,22 = 10.97, P = 0.0032, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial #4, P = 0.0026, trial #5, P = 0.0011, trial #6, P = 0.0277. D2 Re: P = 0.0052, unpaired two-sided t -test. CNO(hM3Dq), n = 12 mice; Saline, n = 12 mice. j CNO(hM4Di) vs Saline, D1 TFC, F 1, 19 = 6.323, P = 0.0211, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial #4, P = 0.0194. D2 Re: P = 0.012, unpaired two-sided t -test. CNO(hM4Di), n = 10 mice; Saline, n = 11 mice). Statistics as indicated, ** P < 0.01, * P < 0.05. Source data are provided as a Source Data file.

Journal: Nature Communications

Article Title: Prefrontal chandelier cells encode stimulus salience to influence learning in male mice

doi: 10.1038/s41467-026-68959-3

Figure Lengend Snippet: a Viral injection, fiber implantation, and representative NpHR expression in mPFC ChCs. b , c Behavioral effects of ChC inhibition during conditioning, applied as indicated (NpHR group (orange) vs EYFP group (black): D1 TFC, F 1,17 = 21.57, P = 0.0002, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial #3, P = 0.0087, trial #4, P = 0.0052, trial #5, P < 0.0001, trial #6, P = 0.0348. D2 Re, NpHR vs EYFP, P = 0.0047, unpaired two-sided t -test. NpHR, n = 11 mice; EYFP, n = 8 mice). c D1 TFC, F 1,16 = 11.69, P = 0.0035, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial #3, P = 0.0495, trial #6, P = 0.0153. D2 Re, NpHR vs EYFP, P = 0.0038, unpaired two-sided t -test. NpHR, n = 9 mice; EYFP, n = 9 mice). d , e Schematics of AAV injections and experimental design for chemogenetic manipulation of ChC activity, with representative images of GCaMP7s co-expressed with hM3Dq ( n = 7 mice) or hM4Di ( n = 9 mice). f , g Corresponding average traces and quantification of ChC Ca²⁺ responses to tone stimuli at baseline and after CNO ( f hM3Dq, P = 0.0029, paired two-sided t -test, n = 7 mice. g hM4Di, P = 0.0075, paired two-sided t -test, n = 9 mice). h Schematics and representative hM3Dq expression in mPFC ChCs. i , j Behavioral effects of chemogenetic activation or inhibition of ChCs during conditioning ( i CNO(hM3Dq) vs Saline, D1 TFC, F 1,22 = 10.97, P = 0.0032, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial #4, P = 0.0026, trial #5, P = 0.0011, trial #6, P = 0.0277. D2 Re: P = 0.0052, unpaired two-sided t -test. CNO(hM3Dq), n = 12 mice; Saline, n = 12 mice. j CNO(hM4Di) vs Saline, D1 TFC, F 1, 19 = 6.323, P = 0.0211, two-way repeated-measures ANOVA; Sidak’s multiple comparison test (two-sided): trial #4, P = 0.0194. D2 Re: P = 0.012, unpaired two-sided t -test. CNO(hM4Di), n = 10 mice; Saline, n = 11 mice). Statistics as indicated, ** P < 0.01, * P < 0.05. Source data are provided as a Source Data file.

Article Snippet: Neutral tone (8 kHz, 60 dB, 2 s) and electric foot shock (0.6 mA, 2 s) were delivered by the standard fear conditioning shock box (UGO BASILE SRL, Italy).

Techniques: Injection, Expressing, Inhibition, Comparison, Activity Assay, Activation Assay, Saline

Comparison of contextual and cued fear conditioning responses between male and female WT and iNOS KO mice. (a) Graphical representation of contextual fear conditioning paradigm. (b) Conditioning and extinction over time in contextual fear conditioning. (c) Fear expression in contextual fear conditioning. (d) Extinction retrieval in contextual fear conditioning. (e) Graphical representation of cued fear conditioning paradigm. (f) Conditioning, extinction, and retrieval over time in cued fear conditioning. (g) Fear expression in cued fear conditioning. (h) Total extinction retrieval in cued fear conditioning. Data are presented as Mean ± SEM and were analyzed using repeated-measures ANOVA (b and f), two-way ANOVA (c), or the non-parametric Kruskal-Wallis test (d, g, and h). A total of 10–18 animals per group were used. @p < 0.05 compared between sexes; #p < 0.05, ##p < 0.01, ###p < 0.001 compared between genotypes. *p < 0.05, ****p < 0.0001 for time comparisons.

Journal: bioRxiv

Article Title: Sex-Dependent Vulnerability to PTSD-Like Behaviors in iNOS Knockout Mice

doi: 10.1101/2025.08.27.672399

Figure Lengend Snippet: Comparison of contextual and cued fear conditioning responses between male and female WT and iNOS KO mice. (a) Graphical representation of contextual fear conditioning paradigm. (b) Conditioning and extinction over time in contextual fear conditioning. (c) Fear expression in contextual fear conditioning. (d) Extinction retrieval in contextual fear conditioning. (e) Graphical representation of cued fear conditioning paradigm. (f) Conditioning, extinction, and retrieval over time in cued fear conditioning. (g) Fear expression in cued fear conditioning. (h) Total extinction retrieval in cued fear conditioning. Data are presented as Mean ± SEM and were analyzed using repeated-measures ANOVA (b and f), two-way ANOVA (c), or the non-parametric Kruskal-Wallis test (d, g, and h). A total of 10–18 animals per group were used. @p < 0.05 compared between sexes; #p < 0.05, ##p < 0.01, ###p < 0.001 compared between genotypes. *p < 0.05, ****p < 0.0001 for time comparisons.

Article Snippet: Mice were habituated to a conditioning box (Ugo Basile Fear conditioning system) and subsequently received three inescapable footshocks (0.75 mA; 2s each) through a metal grid (day 1) to assess the behavioral fear response (freezing) ( ).

Techniques: Comparison, Expressing

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