(A) Generation of the α2F/F control and α2CA1KO, α2CA3KO, α2DGKO mice. (B) Top: False color images showing the α2 staining intensity in immunohistochemically stained sections. Cooler colors = less …
(A) False color images showing α2 staining intensity in immunohistochemically stained sections in α2F/F mice.Cooler colors = less staining. (B) Same as A for α2CA1KO, (C) Same as A for α2CA3KO, (D) …
(A) Top: Representative gray-scale images showing the α1 staining intensity in immunohistochemically stained sections from the hippocampus, cortex and the amygdala of α2F/F, α2CA1KO, α2CA3KO and …
(A) Gabra3 expression in CA1, CA3, DG, cortex and the amygdala of α2F/F, α2CA1KO, α2CA3KO and α2DGKO mice expressed as percentage of α2F/F control. (B) Same as A for Gabra4. *p<0.05 in comparison to …
Mean ( ± S.E.M.) decay time, frequency, amplitude of mIPSCs recorded from (A) the CA1 pyramidal neurons in α2CA1KO and α2F/F sections, (B) the CA3 pyramidal neurons in α2CA3KO and α2F/F sections, (C)…
(A) Mean ( ± S.E.M.) percentage of time spent freezing during the tone 24 hr after auditory fear-conditioning using a delay (black) or trace (white) protocol.*p<0.05, ***p<0.001, different from the …
(A) Left: Activity heat maps on the EPM of representative α2F/F, α2CA1KO, α2CA3KO and α2DGKO mice treated with vehicle or diazepam. Right: Percentage (Mean ± S.E.M.) of time spent in the open arms …
Raw data for elevated plus maze and light/dark box figures.
(A) Left: Percentage of entries into the open arms in elevated plus maze in F/F control, α2CA1KO, α2CA3KO and α2DGKO mice bred on the C57BL/6J background. Right: Total distance travelled in the …
(A) Representative recordings of “Startle Stimulus Only” and “Tone + Startle Stimulus” trials in a α2F/F control mouse treated with vehicle. The increased startle amplitude in “Tone + Startle” …
Raw data for fear-potentiated startle and Vogel conflict test figures.
(A-D) Mean (± S.E.M.) startle amplitude in response to different intensities of white noise bursts measured in arbitrary units based on the displacement of load cells during the first three days …
(A) Stimulation and recording sites and representative traces showing vHPC theta activity before and after diazepam injection in a α2F/F mouse. (B-E) Mean ( ± S.E.M.) peak frequency in the theta …
Raw data for peak frequency and stimulation intensity – peak frequency slope figures.
A-D Mean ( ± S.E.M.) peak frequency in the theta range in dHPC recorded before, 30min after and 60min after diazepam administration in (A) α2F/F, (B) α2CA1KO, (C) α2CA3KO, (D) α2DGKO mice.**p<0.01, …
Results of omnibus statistical tests of experiments for the general characterization of α2CA1KO, α2CA3KO and α2DGKO mice.
1. Immunohistochemistry | ||||||
One-Way ANOVA; Factor: Genotype | ||||||
α2 Subunit | α1 Subunit | α5 Subunit | ||||
CA1 | F(3,10)=9.44 | P=0.003 | F(3,9)=0.08 | p=0.97 | F(3,9)=0.10 | p=0.96 |
CA3 | F(3,10)=5.05 | P=0.02 | F(3,9)=0.01 | p=0.99 | F(3,9)=0.13 | p=0.94 |
DG | F(3,10)=9.08 | P=0.003 | F(3,9)=0.00 | p=1.00 | F(3,9)=0.06 | p=0.98 |
Cortex | F(3,10)=1.90 | p=0.19 | F(3,9)=0.01 | p=0.83 | F(3,9)=0.01 | p=0.99 |
Amygdala | F(3,10)=1.28 | p=0.34 | F(3,9)=0.21 | p=0.89 | F(3,9)=0.36 | p=0.78 |
2. Quantitative PCR | ||||||
One-Way ANOVA; Factor: Genotype | ||||||
α2 Subunit | α3 Subunit | α4 Subunit | ||||
CA1 | F(3,16)=10.66 | p<0.001 | F(3,16)=0.10 | p=0.96 | F(3,16)=2.48 | p=0.1 |
CA3 | F(3,16)=10.53 | p<0.001 | F(3,16)=0.67 | p=0.58 | F(3,16)=4.74 | p=0.02 |
DG | F(3,16)=7.32 | p=0.003 | F(3,12)=1.96 | p=0.17 | F(3,12)=2.90 | p=0.08 |
Cortex | F(3,16)=15.69 | p<0.001 | ||||
Amygdala | F(3,16)=3.01 | p=0.06 | ||||
3. Slice Electrophysiology | ||||||
Two-Way Mixed Factorial ANOVA; Factors: Genotype (between-subjects), Drug (within-subjects) | ||||||
CA1 | Amplitude | Frequency | Decay Time | |||
Genotype | F(1,38)=1.83 | p=0.18 | F(1,38)=2.80 | p=0.10 | F(1,38)=1.94 | p=0.17 |
Drug | F(1,38)=1.49 | p=0.23 | F(1,38)=9.38 | p=0.004 | F(1,38)=107.96 | p<0.001 |
Genotype x Drug Interaction | F(1,38)=2.91 | p=0.09 | F(1,38)=3.11 | p=0.08 | F(1,38)=0.82 | p=0.37 |
CA3 | Amplitude | Frequency | Decay Time | |||
Genotype | F(1,21)=2.36 | p=0.14 | F(1,21)=1.85 | p=0.19 | F(1,21)=0.95 | p=0.34 |
Drug | F(1,21)=0.66 | p=0.42 | F(1,21)=0.09 | p=0.77 | F(1,21)=30.54 | p<0.001 |
Genotype x Drug Interaction | F(1,21)=0.21 | p=0.65 | F(1,21)=1.90 | p=0.18 | F(1,21)=0.25 | p=0.62 |
DG | Amplitude | Frequency | Decay Time | |||
Genotype | F(1,21)=1.50 | p=0.23 | F(1,21)=1.58 | p=0.22 | F(1,21)=0.01 | p=0.91 |
Drug | F(1,21)=2.58 | p=0.12 | F(1,21)=1.58 | p=0.22 | F(1,21)=47.42 | p<0.001 |
Genotype x Drug Interaction | F(1,21)=0.54 | p=0.47 | F(1,21)=1.33 | p=0.26 | F(1,21)=0.30 | p=0.59 |
4. Tests of Hippocampal Function | ||||||
Delay – Trace Fear Conditioning | ||||||
Two-Way Factorial ANOVA; Factors: Genotype (between-subjects), Condition (between-subjects) | ||||||
% Freezing | ||||||
Genotype | F(3,49)=0.38 | p=0.77 | ||||
Condition | F(1,49)=41.57 | p<0.001 | ||||
Genotype x Cond. Interaction | F(3,49)=0.71 | p=0.55 | ||||
Contextual Fear Conditioning | ||||||
One-Way ANOVA; Factor: Genotype (between-subjects) | ||||||
% Freezing | ||||||
Genotype | F(3,35)=5.47 | p=0.003 | ||||
Morris Water Maze | ||||||
Two-Way Factorial ANOVA; Factors: Genotype (between-subjects), Day (within-subjects) | ||||||
Time to platform | ||||||
Genotype | F(3,80)=5.17 | p=0.01 | ||||
Day | F(5,80)=244.12 | p<0.001 | ||||
Genotype x Day Interaction | F(5,80)=1.36 | p=0.19 |
Results of omnibus statistical tests of measured parameters in behavioral tests of anxiety and general locomotion.
1. Elevated Plus Maze (C57BL/6J) | ||||||
Two-Way Factorial ANOVA; Factors: Genotype (between-subjects), Drug (between-subjects) | ||||||
% Open Arm Time | % Open Arm Entries | Distance Travelled | ||||
Genotype | F(3,75)=1.41 | p=0.25 | F(3,75)=0.13 | p=0.94 | F(3,75)=0.85 F(3, 69) = 1.11 | p=0.47 |
Drug | F(1,75)=16.48 | p<0.001 | F(3,75)=2.33 | p=0.13 | F(3,75)=0.61 | p=0.44 |
Genotype x Drug Interaction | F(3,63)=1.54 | p=0.21 | F(3,75)=1.47 | p=0.23 | F(3,75)=1.13 | p=0.34 |
1’. Elevated Plus Maze (129X1/SvJ) | ||||||
Two-Way Factorial ANOVA; Factors: Genotype (between-subjects), Drug (between-subjects) | ||||||
% Open Arm Time | % Open Arm Entries | Distance Travelled | ||||
Genotype | F(3,55)=0.17 | p=0.92 | F(3,55)=1.59 | p=0.20 | F(3,55)=1.17 | p=0.33 |
Drug | F(1,55)=11.49 | P=0.001 | F(1,55)=3.4649 | p=0.07 | F(1,55)=1.28 | p=0.29 |
Genotype x Drug Interaction | F(3,55)=2.28 | p=0.09 | F(3,55)=0.50 | p=0.69 | F(3,55)=0.82 | p=0.49 |
2. Light / Dark Box | ||||||
Two-Way Factorial ANOVA; Factors: Genotype (between-subjects), Drug (between-subjects) | ||||||
% Time in Light | Entries to Light | |||||
Genotype | F(3,73)=5.03 | p=0.003 | F(3,73)=0.84 | p=0.48 | ||
Drug | (F(1,73)=26.00 | p<0.001 | F(1,73)=1.45 | p=0.23 | ||
Genotype x Drug Interaction | F(3,73)=5.53 | p=0.002 | F(3,73)=2.17 | p=0.09 | ||
3. Open Field | ||||||
Two-Way Factorial ANOVA; Factors: Genotype (between-subjects), Drug (between-subjects) | ||||||
Distance Travelled | ||||||
Genotype | F(3,56)=1.78 | p=0.16 | ||||
Drug | F(1,56)=0.04 | p=0.84 | ||||
Genotype x Drug Interaction | F(3,56)=0.43 | p=0.73 |
Results of omnibus statistical tests of measured parameters in behavioral tests of fear.
1. Fear-Potentiated Startle | ||||
---|---|---|---|---|
Within-Genotype Comparisons | ||||
Two-Way Factorial ANOVA; Factors: Tone/No Tone (within-subjects), Drug (between-subjects) | ||||
α2F/F | α2CA1KO | |||
Tone | F(1,28)=33.75 | p<0.001 | F(1,20)=49.17 | p<0.001 |
Drug | F(1,28)=0.30 | p=0.59 | F(1,20)=0.51 | p=0.48 |
Tone x Drug Interaction | F(1,28)=9.95 | p=0.004 | F(1,20)=0.02 | P=0.89 |
α2CA3KO | α2DGKO | |||
Tone | F(1,18)=16.60 | p<0.001 | F(1,20)=54.46 | p<0.001 |
Drug | F(1,18)=0.09 | p=0.77 | F(1,20)=0.73 | p=0.40 |
Tone x Drug Interaction | F(1,18)=5.16 | p=0.04 | F(1,20)=10.24 | p=0.01 |
Between-Genotype Comparisons | ||||
Two-Way Factorial ANOVA; Factors: Genotype (between-subjects), Drug (between-subjects) | ||||
% FPS | ||||
Genotype | F(3,86)=1.17 | p=0.91 | ||
Drug | F(1,86)=16.69 | p<0.001 | ||
Genotype x Drug Interaction | F(3,86)=2.44 | p=0.07 | ||
2. Vogel Conflict Test | ||||
Pretest (Unpunished) Drinking | ||||
One-Way ANOVA; Factor: Genotype (between-subjects) | ||||
Number of licks | ||||
Genotype | F(3,63)=0.63 | p=0.60 | ||
Test (Punished) Drinking | ||||
Two-Way Factorial ANOVA; Factors: Genotype (between-subjects), Drug (between-subjects) | ||||
Number of licks | ||||
Genotype | F(3,59)=1.00 | p=0.40 | ||
Drug | F(1,59)=14.57 | p<0.001 | ||
Genotype x Drug Interaction | F(3,59)=1.21 | p=0.31 |
Omnibus statistical tests of measured parameters in in vivo LFP recordings collected from ventral and dorsal hippocampus.
VENTRAL HIPPOCAMPUS | ||||
---|---|---|---|---|
1. Peak Theta Frequency | ||||
Two-Way ANOVA; Factors: Stimulation intensity (within-subjects), Time before/after drug (within-subjects) | ||||
α2F/F | α2CA1KO | |||
Stimulation intensity | F(4,32)=9.13 | p<0.001 | F(4,24)=18.12 | p<0.001 |
Time after drug | F(2,32)=22.98 | p<0.001 | F(2,24)=37.01 | p<0.001 |
Stimulation x Time | F(8, 32)=4.27 | p<0.001 | F(8,24)=9.07 | p<0.001 |
α2CA3KO | α2DGKO | |||
Stimulation intensity | F(4,32)=8.14 | p<0.001 | F(4,32)=3.84 | P=0.02 |
Time after drug | F(2,32)=6.14 | P=0.02 | F(2,32)=2.47 | P=0.15 |
Stimulation x Time | F(8, 32)=3.14 | P=0.01 | F(8, 32)=6.63 | p<0.001 |
2. Stimulation Intensity – Theta Frequency Slope | ||||
One-Way ANOVA; Factor: Genotype (between-subjects) | ||||
30min post-diazepam | 60min post-diazepam | |||
Genotype | F(3,15)=3.94 | p=0.03 | F(3,73)=0.84 | p=0.48 |
3. Normalized Power | ||||
One-Way ANOVA; Factor: Genotype (between-subjects) | ||||
60min post-diazepam | ||||
Genotype | F(3,15)=4.65 | p=0.02 | ||
DORSAL HIPPOCAMPUS | ||||
4. Peak Theta Frequency | ||||
Two-Way ANOVA; Factors: Stimulation intensity (within-subjects), Time before/after drug (within-subjects) | ||||
α2F/F | α2CA1KO | |||
Stimulation intensity | F(4,32)=8.19 | p<0.001 | F(4,32)=5.29 | p=0.01 |
Time after drug | F(2,32)=31.65 | p<0.001 | F(2,32)=3.03 | p<0.11 |
Stimulation x Time | F(8,32)=8.87 | p=0.003 | F(8,32)=1.89 | p=0.10 |
α2CA3KO | α2DGKO | |||
Stimulation intensity | F(4,40)=19.19 | p<0.001 | F(4,32)=8.86 | p<0.001 |
Time after drug | F(2,40)=27.08 | p<0.001 | F(2,32)=9.25 | p=0.01 |
Stimulation x Time | F(8,40)=4.30 | p<0.001 | F(8,32)=2.39 | p=0.04 |
5. Stimulation Intensity – Theta Frequency Slope | ||||
One-Way ANOVA; Factor: Genotype (between-subjects) | ||||
30min post-diazepam | 60min post-diazepam | |||
Genotype | F(3,17)=2.97 | p=0.14 | F(3,17)=4.32 | p=0.02 |
6. Normalized Power | ||||
One-Way ANOVA; Factor: Genotype (between-subjects) | ||||
60min post-diazepam | ||||
Genotype | F(3,15)=1.02 | p=0.41 |