G2C::Genetics

PSD-95-PDZ3 knock-out mouse

S.G.N. Grant and the G2C Consortium

Corresponding email: Seth.Grant@ed.ac.uk  

 

G2CMine Data Warehouse

Dlg4 @ G2CMine

Genetic and Genomic Information

Gene symbol Dlg4
MGI ID MGI:1277959
G2Cdb mouse G00000004
Ensembl mouse ENSMUSG00000020886
G2Cdb human G00000029
Ensembl human ENSG00000132535

G2CMine Data Warehouse

G2CMine integrates the scientific findings of the Genes to Cognition Programme that utilised neuroproteomics, psychiatric genetics, high-throughput mouse gene targeting combined with behavioural and electrophysiological phenotyping and informatics in order to develop a general strategy for understanding cognition at the molecular, cellular and systems neuroscience levels.

G2CMine provides comprehensive Gene Ontology, Mammalian Phenotype Ontology, Human Phenotype Ontology, UniProt, genetic and protein interactions, and regional mouse brain expression results, together with the phenotyping results of the G2C Programme.

Mutation

Generation of PSD-95-PDZ3 mutant mice described in Migaud et al (1998)

Genotyping

Dlg4 genotyping described in Migaud et al (1998)

Expression

Dlg4 expression described in Migaud et al (1998)

Breeding

Birth of PSD-95-PDZ3-/- mice did not follow Mendelian ratios with 15% of offspring being homozygous knockouts. Genotypes of 3-week-old pups from PSD-95-PDZ3+/- intercrosses identified 116 wt, 209 PSD-95-PDZ3+/-, and 58 PSD-95-PDZ3-/- progeny (Χ2 p < 0.001). Male and female PSD-95-PDZ3-/- mice developed normally to adulthood and exhibited no gross abnormalities, but showed reduced locomotion and small body size. Backcrosses onto the C57BL/6J background were used to maintain the colony and to generate homozygous and wildtype mice to study.

Overview

Mutant mice showed large overall behavioural difference from wildtypes, with 11 of 16 behaviour variables significantly impacted by this mutation. In the elevated plus maze task, four behavioural variables of five were significantly impacted in mutants. In the open field/novel object exploration task, two behavioural variables of two were significantly impacted in mutants. In the rotarod task, two behavioural variables of three were significantly impacted in mutants. In the fear learning task, one behavioural variable, fear learning trial effect, was significantly increased in mutants. In the contextual memory task, one behavioural variable, contextual memory mean, was significantly decreased in mutants. In the cued memory task, one behavioural variable, cued memory change, was significantly decreased in mutants. Definitions of variables may be found below.

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The G2CMine data warehouse provides cohort summaries and individual mouse observations from the PSD-95-PDZ3 knock-out line phenotyping.

 

Variables shown are: EPM total distance, Total distance (cm) travelled in any arm or central zone of the EPM. EPM max speed, Maximum speed (cm/s) travelled in any arm or central zone of the EPM. EPM % time in open, Percentage of time in the open or closed arms of the EPM spent in open arms. EPM time in centre, Total time (s) spent in the central zone of the EPM. EPM max speed, open vs closed, Difference between the maximum speed (cm/s) observed in the open arms and the closed arms of the EPM. OF, NOE total distance, Total distance travelled (log₁₀ cm) during initial exposure to the open field and in presence of the novel object. NOE vs OF distance travelled, Difference in distance travelled (cm) in presence of the novel object and during initial exposure to open field. RR naive fall time, Fall time on accelerating rotarod (log₁₀ s), naive performance in session 1. RR learning, Learning on rotarod, measured as increase in fall time per trial (s/trial) in session 1. RR memory, Memory on rotarod, measured as excess fall time at middle of session 2 relative to middle of session 1. Fear learning, trial effect, Fear learning, measured as extra % time freezing before third trial compared to % time freezing before first trial. Fear learning, tone effect, Fear learning, measured as increase in % time freezing due to third tone compared to increase in % time freezing due to first tone. Contextual memory, mean, Contextual memory, measured as difference in % time freezing during first 120 s re-exposure to the box compared to first 120 s in the box on previous day. Contextual memory, change, Contextual memory, measured as increase in % time spent freezing from first time bin of 30 s to fourth bin of 30 s during 120 s re-exposure to the box. Cued memory, mean, Cued memory, measured as increase in % time spent freezing during 120 s of tone re-exposure compared to increase in % time spent freezing during initial tone on previous day. Cued memory, change, Cued memory, measured as increase in % time spent freezing from first time bin of 30 s to fourth bin of 30 s during 120 s re-exposure to the tone.

 

Variable Units Wildtype M (n=16) Wildtype F (n=11) Mutant M (n=16) Mutant F (n=13) P(sex×mutation) P(mutation)
EPM total distance cm 1176 (60) 1287 (99) 1003 (106) 1086 (83) 0.87 0.042 *
EPM max speed cm/s 25 (1) 27.1 (2.2) 22 (1.9) 22.5 (1.1) 0.63 0.025 *
EPM % time in open % 22.4 (4) 21.1 (8.2) 53.4 (8.1) 66.5 (6.4) 0.3 0.0000012 ***
EPM time in centre s 56.6 (5.3) 58.2 (6.9) 31.3 (6.7) 42.3 (5.5) 0.45 0.0011 **
EPM max speed, open vs closed cm/s -4.6 (1.3) -5.5 (2.8) -2.9 (2.4) -0.1 (2) 0.39 0.13
OF, NOE total distance log10 cm 3.63 (0.03) 3.74 (0.02) 3.46 (0.05) 3.4 (0.04) 0.038 * 0.00000012 ***
NOE vs OF distance travelled cm -1270 (170) -1150 (230) -830 (130) -760 (120) 0.88 0.012 *
RR naive fall time log10 s 1.41 (0.1) 1.5 (0.12) 1.05 (0.05) 1.35 (0.06) 0.24 0.0024 **
RR learning s/trial 7.5 (1.2) 7.7 (1.7) 2.4 (0.5) 2.6 (1) 0.99 0.000018 ***
RR memory s 17.7 (5) 22.6 (7.5) 10.3 (4.5) 15.4 (4.9) 0.98 0.18
Fear learning, trial effect % freezing 43.4 (5.7) 39 (7.5) 67.6 (5.9) 76.4 (5.2) 0.29 0.0000089 ***
Fear learning, tone effect % freezing 6.2 (3.6) 1.5 (5.9) 2.3 (5.8) -7.1 (10.1) 0.72 0.37
Contextual memory, mean % freezing 38.5 (4.8) 29 (5.8) 23.1 (3.9) 13.8 (5.7) 0.99 0.0033 **
Contextual memory, change % freezing 14.7 (5.3) -4.1 (11.9) 2 (6.2) -18 (7.6) 0.94 0.085
Cued memory, mean % freezing -1.1 (4.6) 10 (8.1) -7.7 (6) -1.7 (11.5) 0.74 0.25
Cued memory, change % freezing 0.7 (7.2) 10.1 (6.8) -10 (5.5) -9 (5.8) 0.53 0.031 *

Elevated Plus Maze

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Variables shown are: EPM total distance, Total distance (cm) travelled in any arm or central zone of the EPM. EPM max speed, Maximum speed (cm/s) travelled in any arm or central zone of the EPM. EPM % time in open, Percentage of time in the open or closed arms of the EPM spent in open arms. EPM time in centre, Total time (s) spent in the central zone of the EPM. EPM max speed, open vs closed, Difference between the maximum speed (cm/s) observed in the open arms and the closed arms of the EPM.

 

Variable Units Wildtype M (n=16) Wildtype F (n=11) Mutant M (n=16) Mutant F (n=13) P(sex×mutation) P(mutation)
EPM total distance cm 1176 (60) 1287 (99) 1003 (106) 1086 (83) 0.87 0.042 *
EPM max speed cm/s 25 (1) 27.1 (2.2) 22 (1.9) 22.5 (1.1) 0.63 0.025 *
EPM % time in open % 22.4 (4) 21.1 (8.2) 53.4 (8.1) 66.5 (6.4) 0.3 0.0000012 ***
EPM time in centre s 56.6 (5.3) 58.2 (6.9) 31.3 (6.7) 42.3 (5.5) 0.45 0.0011 **
EPM max speed, open vs closed cm/s -4.6 (1.3) -5.5 (2.8) -2.9 (2.4) -0.1 (2) 0.39 0.13

Open Field/Novel Object

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Variables shown are: OF, NOE total distance, Total distance travelled (log₁₀ cm) during initial exposure to the open field and in presence of the novel object. NOE vs OF distance travelled, Difference in distance travelled (cm) in presence of the novel object and during initial exposure to open field.

 

Variable Units Wildtype M (n=16) Wildtype F (n=11) Mutant M (n=16) Mutant F (n=13) P(sex×mutation) P(mutation)
OF, NOE total distance log10 cm 3.63 (0.03) 3.74 (0.02) 3.46 (0.05) 3.4 (0.04) 0.038 * 0.00000012 ***
NOE vs OF distance travelled cm -1270 (170) -1150 (230) -830 (130) -760 (120) 0.88 0.012 *

Rotarod

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Variables shown are: RR naive fall time, Fall time on accelerating rotarod (log₁₀ s), naive performance in session 1. RR learning, Learning on rotarod, measured as increase in fall time per trial (s/trial) in session 1. RR memory, Memory on rotarod, measured as excess fall time at middle of session 2 relative to middle of session 1.

 

Variable Units Wildtype M (n=16) Wildtype F (n=11) Mutant M (n=16) Mutant F (n=13) P(sex×mutation) P(mutation)
RR naive fall time log10 s 1.41 (0.1) 1.5 (0.12) 1.05 (0.05) 1.35 (0.06) 0.24 0.0024 **
RR learning s/trial 7.5 (1.2) 7.7 (1.7) 2.4 (0.5) 2.6 (1) 0.99 0.000018 ***
RR memory s 17.7 (5) 22.6 (7.5) 10.3 (4.5) 15.4 (4.9) 0.98 0.18

Fear Conditioning

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Variables shown are: Fear learning, trial effect, Fear learning, measured as extra % time freezing before third trial compared to % time freezing before first trial. Fear learning, tone effect, Fear learning, measured as increase in % time freezing due to third tone compared to increase in % time freezing due to first tone. Contextual memory, mean, Contextual memory, measured as difference in % time freezing during first 120 s re-exposure to the box compared to first 120 s in the box on previous day. Contextual memory, change, Contextual memory, measured as increase in % time spent freezing from first time bin of 30 s to fourth bin of 30 s during 120 s re-exposure to the box. Cued memory, mean, Cued memory, measured as increase in % time spent freezing during 120 s of tone re-exposure compared to increase in % time spent freezing during initial tone on previous day. Cued memory, change, Cued memory, measured as increase in % time spent freezing from first time bin of 30 s to fourth bin of 30 s during 120 s re-exposure to the tone.

 

Variable Units Wildtype M (n=16) Wildtype F (n=11) Mutant M (n=16) Mutant F (n=13) P(sex×mutation) P(mutation)
Fear learning, trial effect % freezing 43.4 (5.7) 39 (7.5) 67.6 (5.9) 76.4 (5.2) 0.29 0.0000089 ***
Fear learning, tone effect % freezing 6.2 (3.6) 1.5 (5.9) 2.3 (5.8) -7.1 (10.1) 0.72 0.37
Contextual memory, mean % freezing 38.5 (4.8) 29 (5.8) 23.1 (3.9) 13.8 (5.7) 0.99 0.0033 **
Contextual memory, change % freezing 14.7 (5.3) -4.1 (11.9) 2 (6.2) -18 (7.6) 0.94 0.085
Cued memory, mean % freezing -1.1 (4.6) 10 (8.1) -7.7 (6) -1.7 (11.5) 0.74 0.25
Cued memory, change % freezing 0.7 (7.2) 10.1 (6.8) -10 (5.5) -9 (5.8) 0.53 0.031 *

Overview

Mutant slices showed profound overall electrophysiological difference from wildtype slices, with eight of nine variables significantly impacted in these mutant slices. In the basal synaptic transmission experiment, max fEPSP amplitude was significantly decreased in mutants. In the theta burst stimulation experiment, five electrophysiological variables of five were significantly impacted in mutants. In the long term potentiation experiment, two electrophysiological variables of two were significantly impacted in mutants. Only the paired pulse facilitation experiment was unaffected. Note that in the theta burst analysis, the tenth burst is analysed as representative of individual burst amplitude. Definitions of variables may be found below.

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The G2CMine data warehouse provides slice group summaries and individual mouse observations from the PSD-95-PDZ3 knock-out line phenotyping.

 

Variables shown are: Max fEPSP amplitude, Maximum field excitatory postsynaptic potential (fEPSP) amplitude. PPF at 50ms, ampl ratio, Paired pulse facilitation (PPF), pulses separated by 50ms, amplitude ratio. Burst 1, PPF at 10ms, ampl ratio, Paired pulse facilitation (PPF), pulses separated by 10ms, amplitude ratio, observed during first two pulses of the first 100Hz burst during theta-burst stimulation. Burst 1, EPSP3 depr, ampl ratio, Depression observed in third fEPSP relative to the second fEPSP of the first 100Hz burst, amplitude ratio. Burst 1, ave amplitude, Average amplitude of four fEPSPs in first burst. Burst 2, ave amplitude, Average amplitude of four fEPSPs in second burst. Burst 3, ave amplitude, Average amplitude of four fEPSPs in third burst. Burst 4, ave amplitude, Average amplitude of four fEPSPs in fourth burst. Burst 5, ave amplitude, Average amplitude of four fEPSPs in fifth burst. Burst 6, ave amplitude, Average amplitude of four fEPSPs in sixth burst. Burst 7, ave amplitude, Average amplitude of four fEPSPs in seventh burst. Burst 8, ave amplitude, Average amplitude of four fEPSPs in eighth burst. Burst 9, ave amplitude, Average amplitude of four fEPSPs in ninth burst. Burst 10, ave amplitude, Average amplitude of four fEPSPs in tenth burst. Burst 2-4 ave amplitude vs Burst 1, Facilitation observed in average amplitude of bursts 2-4, relative to average amplitude of burst 1. Burst 8-10 ave amplitude vs Burst 2-4, Depression observed in average amplitude of bursts 8-10, relative to average amplitude of bursts 2-4. LTP vs PTP, amplitude ratio, Reduction in potentiation from immediately after theta-burst stimulation to one hour later, fEPSP amplitude ratio. LTP based on amplitude, Long term potentiation, ratio of amplitudes of fEPSPs in test pathway and control pathway.

 

Variable Units Wildtype slices (animals) Wildtype mean (SEM) Mutant slices (animals) Mutant mean (SEM) P(animals) P(mutation)
Max fEPSP amplitude µV 14 (4) 3249 (190) 20 (5) 1240 (67) 0.52 1e-05 ***
PPF at 50ms, ampl ratio % 14 (4) 178.9 (5.3) 20 (5) 202.9 (6.7) 0.2 0.063
Burst 1, PPF at 10ms, ampl ratio % 14 (4) 190.2 (7) 20 (5) 228.3 (5.2) 0.3 0.0052 **
Burst 1, EPSP3 depr, ampl ratio % 14 (4) 65.8 (2.8) 20 (5) 109.9 (2) 0.73 1.5e-06 ***
Burst 1, ave amplitude µV 14 (4) 1331 (84) 20 (5) 381 (26) 0.17 2.1e-05 ***
Burst 2, ave amplitude µV 14 (4) 2231 (148) 20 (5) 1027 (58) 0.92 7.6e-06 ***
Burst 3, ave amplitude µV 14 (4) 2463 (160) 20 (5) 1232 (69) 0.94 9.5e-06 ***
Burst 4, ave amplitude µV 14 (4) 2494 (160) 20 (5) 1364 (76) 0.95 1.8e-05 ***
Burst 5, ave amplitude µV 14 (4) 2443 (154) 20 (5) 1404 (75) 0.96 1.9e-05 ***
Burst 6, ave amplitude µV 14 (4) 2381 (149) 20 (5) 1400 (75) 0.93 3.6e-05 ***
Burst 7, ave amplitude µV 14 (4) 2297 (141) 20 (5) 1367 (73) 0.96 2.8e-05 ***
Burst 8, ave amplitude µV 14 (4) 2220 (135) 20 (5) 1326 (71) 0.95 3.1e-05 ***
Burst 9, ave amplitude µV 14 (4) 2155 (132) 20 (5) 1288 (71) 0.96 2.9e-05 ***
Burst 10, ave amplitude µV 14 (4) 2106 (130) 20 (5) 1246 (69) 0.96 2.9e-05 ***
Burst 2-4 ave amplitude vs Burst 1 % 14 (4) 181.2 (8.3) 20 (5) 237.8 (7) 0.36 0.0019 **
Burst 8-10 ave amplitude vs Burst 2-4 % 14 (4) 90.6 (1.4) 20 (5) 103.6 (1.4) 0.64 0.00027 ***
LTP vs PTP, amplitude ratio % 14 (4) 80.8 (1) 20 (5) 69.4 (1.6) 0.48 0.0011 **
LTP based on amplitude % 14 (4) 173.9 (6.5) 20 (5) 222.7 (10.8) 0.3 0.017 *

Basal Synaptic Transmission

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Variables shown are: Max fEPSP amplitude, Maximum field excitatory postsynaptic potential (fEPSP) amplitude.

 

Variable Units Wildtype slices (animals) Wildtype mean (SEM) Mutant slices (animals) Mutant mean (SEM) P(animals) P(mutation)
Max fEPSP amplitude µV 14 (4) 3249 (190) 20 (5) 1240 (67) 0.52 1e-05 ***

Paired Pulse Facilitation

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Variables shown are: PPF at 50ms, ampl ratio, Paired pulse facilitation (PPF), pulses separated by 50ms, amplitude ratio.

 

Variable Units Wildtype slices (animals) Wildtype mean (SEM) Mutant slices (animals) Mutant mean (SEM) P(animals) P(mutation)
PPF at 50ms, ampl ratio % 14 (4) 178.9 (5.3) 20 (5) 202.9 (6.7) 0.2 0.063

Theta Burst Stimulation

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Variables shown are: Burst 1, PPF at 10ms, ampl ratio, Paired pulse facilitation (PPF), pulses separated by 10ms, amplitude ratio, observed during first two pulses of the first 100Hz burst during theta-burst stimulation. Burst 1, EPSP3 depr, ampl ratio, Depression observed in third fEPSP relative to the second fEPSP of the first 100Hz burst, amplitude ratio. Burst 1, ave amplitude, Average amplitude of four fEPSPs in first burst. Burst 2, ave amplitude, Average amplitude of four fEPSPs in second burst. Burst 3, ave amplitude, Average amplitude of four fEPSPs in third burst. Burst 4, ave amplitude, Average amplitude of four fEPSPs in fourth burst. Burst 5, ave amplitude, Average amplitude of four fEPSPs in fifth burst. Burst 6, ave amplitude, Average amplitude of four fEPSPs in sixth burst. Burst 7, ave amplitude, Average amplitude of four fEPSPs in seventh burst. Burst 8, ave amplitude, Average amplitude of four fEPSPs in eighth burst. Burst 9, ave amplitude, Average amplitude of four fEPSPs in ninth burst. Burst 10, ave amplitude, Average amplitude of four fEPSPs in tenth burst. Burst 2-4 ave amplitude vs Burst 1, Facilitation observed in average amplitude of bursts 2-4, relative to average amplitude of burst 1. Burst 8-10 ave amplitude vs Burst 2-4, Depression observed in average amplitude of bursts 8-10, relative to average amplitude of bursts 2-4.

 

Variable Units Wildtype slices (animals) Wildtype mean (SEM) Mutant slices (animals) Mutant mean (SEM) P(animals) P(mutation)
Burst 1, PPF at 10ms, ampl ratio % 14 (4) 190.2 (7) 20 (5) 228.3 (5.2) 0.3 0.0052 **
Burst 1, EPSP3 depr, ampl ratio % 14 (4) 65.8 (2.8) 20 (5) 109.9 (2) 0.73 1.5e-06 ***
Burst 1, ave amplitude µV 14 (4) 1331 (84) 20 (5) 381 (26) 0.17 2.1e-05 ***
Burst 2, ave amplitude µV 14 (4) 2231 (148) 20 (5) 1027 (58) 0.92 7.6e-06 ***
Burst 3, ave amplitude µV 14 (4) 2463 (160) 20 (5) 1232 (69) 0.94 9.5e-06 ***
Burst 4, ave amplitude µV 14 (4) 2494 (160) 20 (5) 1364 (76) 0.95 1.8e-05 ***
Burst 5, ave amplitude µV 14 (4) 2443 (154) 20 (5) 1404 (75) 0.96 1.9e-05 ***
Burst 6, ave amplitude µV 14 (4) 2381 (149) 20 (5) 1400 (75) 0.93 3.6e-05 ***
Burst 7, ave amplitude µV 14 (4) 2297 (141) 20 (5) 1367 (73) 0.96 2.8e-05 ***
Burst 8, ave amplitude µV 14 (4) 2220 (135) 20 (5) 1326 (71) 0.95 3.1e-05 ***
Burst 9, ave amplitude µV 14 (4) 2155 (132) 20 (5) 1288 (71) 0.96 2.9e-05 ***
Burst 10, ave amplitude µV 14 (4) 2106 (130) 20 (5) 1246 (69) 0.96 2.9e-05 ***
Burst 2-4 ave amplitude vs Burst 1 % 14 (4) 181.2 (8.3) 20 (5) 237.8 (7) 0.36 0.0019 **
Burst 8-10 ave amplitude vs Burst 2-4 % 14 (4) 90.6 (1.4) 20 (5) 103.6 (1.4) 0.64 0.00027 ***

Long Term Potentiation

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Variables shown are: LTP vs PTP, amplitude ratio, Reduction in potentiation from immediately after theta-burst stimulation to one hour later, fEPSP amplitude ratio. LTP based on amplitude, Long term potentiation, ratio of amplitudes of fEPSPs in test pathway and control pathway.

 

Variable Units Wildtype slices (animals) Wildtype mean (SEM) Mutant slices (animals) Mutant mean (SEM) P(animals) P(mutation)
LTP vs PTP, amplitude ratio % 14 (4) 80.8 (1) 20 (5) 69.4 (1.6) 0.48 0.0011 **
LTP based on amplitude % 14 (4) 173.9 (6.5) 20 (5) 222.7 (10.8) 0.3 0.017 *
© G2C 2014. The Genes to Cognition Programme received funding from The Wellcome Trust and the EU FP7 Framework Programmes:
EUROSPIN (FP7-HEALTH-241498), SynSys (FP7-HEALTH-242167) and GENCODYS (FP7-HEALTH-241995).

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