Journal: EMBO Molecular Medicine

Article Title: Spironolactone is an antagonist of NRG 1‐ ERBB 4 signaling and schizophrenia‐relevant endophenotypes in mice

doi: 10.15252/emmm.201707691

Figure Lengend Snippet: Experimental design to test PPI deficits in Nrg1 ‐tg mice. Nrg1 ‐tg mice and wt controls were tested in the PPI test 1 day prior to chronic spironolactone treatment. One day after the last injection, transgenic and control mice were submitted to the PPI paradigm. Naïve Nrg1 ‐tg displayed strong PPI deficits prior to chronic spironolactone treatment (effect of genotype F 1,33 = 25.74; P < 0.0001, two‐way ANOVA, and Bonferroni post hoc test: *** P < 0.0001, *** P = 0.0002, *** P = 0.0003 for prepulse intensities 70, 75 and 80 dB, respectively). Chronic treatment with spironolactone improved PPI in transgenic animals (effect of treatment F 1,20 = 9.63; P = 0.0056, two‐way ANOVA), with the most prominent effect when a prepulse of 70 dB was presented (** P = 0.0060, Bonferroni test; 75 and 80 dB were P = 0.3067 and P = 0.1077, respectively, Bonferroni test). Spironolactone treatment did not influenced PPI in wt controls (effect of treatment F 1,26 = 0.14; P = 0.7096, two‐way ANOVA). Nrg‐ tg mice spent less time in the center of the open‐field arena (effect of genotype F 1,44 = 6.69; P = 0.0131, two‐way ANOVA). Spironolactone treatment did not influence this parameter ( F 1,44 = 0.12; P = 0.7290, two‐way ANOVA). In the open‐field test, Nrg1 ‐tg mice showed increased defecation (effect of genotype F 1,44 = 10.35; P = 0.0024, two‐way ANOVA), which was significant in vehicle‐treated animals (* P = 0.0310; Bonferroni test), but not in spironolactone‐treated animals ( P = 0.0967). There was no effect of treatment observed ( F 1,44 = 0.65; P = 0.4256, two‐way ANOVA). Transgenic animals displayed more frequent urination than wt controls (effect of genotype F 1,44 = 14.14; P = 0.0005, two‐way ANOVA) in both vehicle and spironolactone‐treated animals (* P = 0.0451 for vehicle‐treated and * P = 0.0100 for spironolactone‐treated groups, Bonferroni test), without effect of treatment ( F 1,44 = 0.17; P = 0.6781, two‐way ANOVA). In the light–dark preference test, transgenic mice performed similar number of transitions between light and dark compartment when compared to wt controls (effect of genotype F 1,41 = 0.05; P = 0.8227, two‐way ANOVA). In the tail suspension test, transgenic mice were more active than wt controls. The effect was independent of treatment (effect of genotype F 1,44 = 35.16; P < 0.0001, two‐way ANOVA). The Bonferroni test showed a significant difference between genotypes both in vehicle and spironolactone‐treated groups (*** P < 0.0001 and ** P = 0.0067, respectively). Nrg1 ‐tg animals displayed an increased activity in the Y‐maze test as they performed more arm choices (effect of genotype F 1,44 = 14.56; P = 0.0004, two‐way ANOVA), with * P = 0.0301 in vehicle‐treated and * P = 0.0127 in spironolactone‐treated groups (Bonferroni test). There was no effect of treatment ( F 1,44 = 0.39; P = 0.5367, two‐way ANOVA). Naïve Nrg1 ‐tg mice did not display impairments in the fear conditioning test. However, transgenic animals treated with spironolactone showed a tendency toward lower freezing rate during contextual memory test ( P = 0.0885; Mann–Whitney test). A two‐way ANOVA yielded a substantial, but not significant, interaction in the contextual memory test ( F 1,44 = 3.56: P = 0.0657); but neither an effect of genotype ( F 1,44 = 0.96; P = 0.3315) nor of treatment ( F 1,44 = 1.16; P = 0.2880) was detected. Naïve Nrg1 ‐tg showed an increased freezing rate during baseline recordings prior to the cue test ( ## P = 0.0351, Mann–Whitney test); this difference between the genotypes was not observed in groups treated with spironolactone. In the hot plate test, Nrg1 ‐tg mice showed an increased latency to the first reaction (effect of genotype F 1,44 = 12.47: P = 0.0010; two‐way ANOVA), which was most prominent in the group treated with spironolactone (** P = 0.0024, Bonferroni test) but not in vehicle‐treated groups ( P = 0.2662, Bonferroni test). Spironolactone treatment did not influence PPI in wt mice (effect of treatment F 1,22 = 0.00; P = 0.9461, two‐way ANOVA). Data information: Data are shown as mean, and error bars represent SEM; n.s., not significant; Spiro, spironolactone; pre‐Spiro, data collected before spironolactone treatment. n = 12 per genotype and treatment with an exception in (F): Nrg1‐ tg vehicle, n = 11; Nrg1‐ tg Spiro, n = 10; wt vehicle, n = 12; wt Spiro, n = 12).

Article Snippet: Fear memory assessment that is measured by freezing behavior was performed using the Ugo Basile Fear Conditioning System (Varese, Italy).

Techniques: Injection, Transgenic Assay, Control, Suspension, Activity Assay, MANN-WHITNEY, Hot Plate Test

Journal: Nature Communications

Article Title: Experience-dependent resonance in amygdalo-cortical circuits supports fear memory retrieval following extinction

doi: 10.1038/s41467-020-18199-w

Figure Lengend Snippet: a Firing rate of BLA MUA is not significantly different during 4 Hz and 8 Hz sinusoidal stimulation in the conditioned context (firing rate normalized to no light; paired two-tailed Wilcoxon test: P = 0.22, n = 50 MUA). b Same as in a but in unconditioned context (paired two-tailed Wilcoxon test: P = 0.33, n = 50 MUA). c Averaged PPC spectra (left) of BLA MUA during 4 Hz and 8 Hz optical stimulation in the conditioned context (Fear + Ext memory state). MUA phase-locking quantified as 4:8 Hz PPC ratio (right) is significantly shifted by 4 Hz vs. 8 Hz stimulation in the memory-context (paired two-tailed Wilcoxon test: P = 0.0005, n = 50 MUA). d Same as in c but in the unconditioned context (no memory state). MUA phase-locking is not significantly shifted by 4 Hz vs. 8 Hz stimulation in the unconditioned context (paired two-tailed Wilcoxon test: P = 0.17, n = 50 MUA). e Venn diagram showing the distribution of units significantly phase-locked to 4 Hz stimulus waveform in the conditioned (Fear + Ext) context (left), unconditioned neutral context (right), or both (middle overlap). f Proportion of units recruited to freezing-associated 4 Hz oscillations in the conditioned context is enriched among units that significantly phase-lock to 4 Hz stimulation in the conditioned context (Chi-square test: X 2 = 8.24, P = 0.0041, n = 17 units). g Same as e but for 8 Hz stimulation. h Proportion of units recruited to non-freezing-associated 8 Hz oscillations in the conditioned context among units that significantly phase-lock to 8 Hz stimulation in the conditioned versus unconditioned context (Chi-square test: X 2 = 1.94, P = 0.16, n = 14 units: one of the six Fear+Ext-only units in panel g did not meet the minimum number of spike criterium for PPC analysis during the non-freezing behavioral state). All error bars and shaded area: mean ± SEM.

Article Snippet: MUA phase-locking is not significantly shifted by 4 Hz vs. 8 Hz stimulation in the unconditioned context (paired two-tailed Wilcoxon test: P = 0.17, n = 50 MUA). e Venn diagram showing the distribution of units significantly phase-locked to 4 Hz stimulus waveform in the conditioned (Fear + Ext) context (left), unconditioned neutral context (right), or both (middle overlap). f Proportion of units recruited to freezing-associated 4 Hz oscillations in the conditioned context is enriched among units that significantly phase-lock to 4 Hz stimulation in the conditioned context (Chi-square test: X = 8.24, P = 0.0041, n = 17 units). g Same as e but for 8 Hz stimulation. h Proportion of units recruited to non-freezing-associated 8 Hz oscillations in the conditioned context among units that significantly phase-lock to 8 Hz stimulation in the conditioned versus unconditioned context (Chi-square test: X = 1.94, P = 0.16, n = 14 units: one of the six Fear+Ext-only units in panel g did not meet the minimum number of spike criterium for PPC analysis during the non-freezing behavioral state).

Techniques: Two Tailed Test

Journal: Nature Communications

Article Title: Experience-dependent resonance in amygdalo-cortical circuits supports fear memory retrieval following extinction

doi: 10.1038/s41467-020-18199-w

Figure Lengend Snippet: a Schematic of optogenetic strategy to increase frequency-specific functional interactions between the BLA and mPFC. b Example images showing ChR2-mCherry expression, with optical fiber and electrode placement in mPFC and BLA. Targeting was similarly confirmed by histological analysis in all 11 mice; see Methods section for detail. Scale bar: 300 µm. c , d Representative cross-power spectrograms illustrating differential effects of 4 Hz in- and anti-phase optical stimulation in the conditioned context ( c ; fear+ext memory state) and unconditioned novel context ( d ; no-memory state). e Averaged cross-power spectra comparing effects of in-phase and anti-phase 4 Hz stimulation to no-stimulation baseline during fear+extinction memory and no-memory states ( n = 8 mice). f Quantification of the cross-power spectra from e . 4 Hz in-phase stimulation increases the 4:8 Hz cross-power ratio compared to the no-stimulation baseline, whereas 4 Hz anti-phase stimulation does not. This effect is absent in a context where the mouse has no fear memory (two-way RM ANOVA: trial F (1,7) = 34.12, P = 0.0006, stimulation F (2,14) = 7.80, P = 0.0053, trial × stimulation F (2,14) = 7.46, P = 0.0062, n = 8 mice. Holm-Sidak’s multiple comparisons test; fear+ext memory state: no light vs. 4 Hz anti-phase: t (14) = 2.01, P = 0.064; no light vs. 4 Hz in-phase t (14) = 3.39, P = 0.0087. No memory state: no light vs. 4 Hz anti-phase: t (14) = 0.33, P = 0.94; no light vs. 4 Hz in-phase t (14) = 0.29, P = 0.94). g 4 Hz in-phase stimulation increases the conditioned freezing response compared to the no-stimulation baseline, whereas 4 Hz anti-phase stimulation does not. This effect is absent in a context where the mouse has no fear memory (two-way repeated measures ANOVA: trial F (1,10) = 15.43, P = 0.0028, stimulation F (2,20) = 10.18, P = 0.0009, trial × stimulation F (2,20) = 5.28, P = 0.014, n = 11 mice. Holm-Sidak’s multiple comparisons test; fear+ext memory state: no light vs. 4 Hz anti-phase: t (20) = 0.55, P = 0.59; no light vs. 4 Hz in-phase t (20) = 5.20, P < 0.0001. No memory state: no light vs. 4 Hz anti-phase: t (20) = 0.0088, P = 0.99; no light vs. 4 Hz in-phase t (20) = 0.97, P = 0.71). All error bars and shaded area: mean ± SEM.

Article Snippet: MUA phase-locking is not significantly shifted by 4 Hz vs. 8 Hz stimulation in the unconditioned context (paired two-tailed Wilcoxon test: P = 0.17, n = 50 MUA). e Venn diagram showing the distribution of units significantly phase-locked to 4 Hz stimulus waveform in the conditioned (Fear + Ext) context (left), unconditioned neutral context (right), or both (middle overlap). f Proportion of units recruited to freezing-associated 4 Hz oscillations in the conditioned context is enriched among units that significantly phase-lock to 4 Hz stimulation in the conditioned context (Chi-square test: X = 8.24, P = 0.0041, n = 17 units). g Same as e but for 8 Hz stimulation. h Proportion of units recruited to non-freezing-associated 8 Hz oscillations in the conditioned context among units that significantly phase-lock to 8 Hz stimulation in the conditioned versus unconditioned context (Chi-square test: X = 1.94, P = 0.16, n = 14 units: one of the six Fear+Ext-only units in panel g did not meet the minimum number of spike criterium for PPC analysis during the non-freezing behavioral state).

Techniques: Functional Assay, Expressing

Journal: Frontiers in Genetics

Article Title: Affective phenotypes in heterozygous LRRK2 R1441G knock-in mice

doi: 10.3389/fgene.2025.1629897

Figure Lengend Snippet: Evidence for depression-related behavioural phenotypes in heterozygous KI +/− mice. (A) All mice showed a preference for the sucrose solution with the percentage of sucrose solution exceeding 50%, but a reduction was evident in the KI +/− group, whose preference was significantly lower than KI +/+ mice ( p < 0.05 *) and marginally below WT mice ( p = 0.06 # ) based on pairwise comparisons by Fisher’s LSD. (B) Latency to shuttle (left) and percentage of escape failures (right) in the conditioned active avoidance task illustrate the impact of prior exposure to inescapable shock versus escapable shock. This difference resulted in a significant main effect of pre-exposure experience (* p < 0.01). (C) Immobility time (as a percentage) across successive 1-min bins on days 1 and 2 are separately depicted, showing earlier and stronger immobility in KI+/− and KI+/+ mice than WT mice (especially on Day 2). (D) Mean immobility time (as a percentage) in the FST is shown for the full 10-min test (bins 1–10, left) and the last 6 min (bins 5–10, right). Both KI+/− and KI+/+ mice exhibited significantly higher immobility than WT mice in post hoc pairwise comparisons ( p < 0.05 *). The SPT and FST were conducted in mice from Cohort A. n = 12 (6♀+ 6♂) per genotype. The LH experiment included mice in both cohorts A and B. n = 12 (6♀+ 6♂) per genotype per pre-exposure condition. All data are presented as the mean ± SE. Overlaid scatter points represent data from individual mice (green dots = ♂; pink dots = ♀).

Article Snippet: Two conditioned freezing chambers (MED-VFC-USB-M, Med Associates, VT, United States) were used.

Techniques:

Journal: Frontiers in Genetics

Article Title: Affective phenotypes in heterozygous LRRK2 R1441G knock-in mice

doi: 10.3389/fgene.2025.1629897

Figure Lengend Snippet: Selected phenotypes in anxiety-related behaviour. (A) The distribution of time spent in the open arms of the EPM relative to time in all arms (in percent) provides a measure of anxiety-like behaviour. A significant difference was detected using Fisher’s LSD test between KI +/− and WT mice (*), while the difference between KI +/− and KI +/+ did not achieve significance (# at p = 0.09). The EPM test included mice in both cohort A and B. n = 24 (12♀+ 12♂) per genotype. Overlaid scatter points represent data from individual mice (green dots = ♂; pink dots = ♀). (B) The expression of conditioned fear to the tone-CS, assessed 48 h after tone–shock pairings, was indexed by percentage time freezing in the tone test. Tone-freezing was continuously tracked for 8 min after the initial 2-min pre-CS period. The data presented are averaged across three consecutive test days (separate plots for each day are presented in ). (C) The expression of foreground contextual fear conditioning was assessed by returning the animals to the shocked context “A” on the first and third days (1A and 3A) and the neutral context “B” on the second and fourth days (2B and 4B) after conditioning. Higher levels of freezing in context “A” relative context “B” supported the expression of context-specific conditioned fear response. For (B,C) , n = 12 (6♀+ 6♂) per genotype. All data are presented as the mean ± SE.

Article Snippet: Two conditioned freezing chambers (MED-VFC-USB-M, Med Associates, VT, United States) were used.

Techniques: Expressing