G2C::Genetics

Mutation

Generation of Dlg3 mutant mice described in Cuthbert et al (2007)

Genotyping

Dlg3 genotyping described in Cuthbert et al (2007)

Expression

Dlg3 expression described in Cuthbert et al (2007)

Breeding

Dlg3 resides on the X chromosome. Birth of SAP102^-/Y mice followed approximately Mendelian ratios with 25% of offspring being hemizygous knockouts. Birth of homozygous knockouts also followed Mendelian ratios with 19% of offspring being homozygous (female) knockouts. Genotypes of 3-week-old pups from heterozygous intercrosses identified 29 male wt, 8 female wt, 34 SAP102^+/-, 42 SAP102^-/Y, and 23 homozygous (female) progeny (Χ² p=0.11). Male SAP102^-/Y and female homozygous mice developed normally to adulthood, exhibited normal body size and no gross abnormalities. Backcrosses onto the 129S5/SvEvBrd background were used to maintain the colony and to generate hemizygous and wildtype mice to study.

Overview

Mutant mice showed large overall behavioural difference from wildtypes, with four of 16 behaviour variables significantly impacted in these mutant mice. In the elevated plus maze task, one behavioural variable, EPM max speed open vs closed, was significantly increased in mutants. In the open field/novel object exploration task, one behavioural variable, OF NOE total distance, was significantly decreased in mutants. In the rotarod task, one behavioural variable, RR naive fall time, was significantly increased in mutants. In the cued memory task, one behavioural variable, cued memory change, was significantly increased in mutants. Only fear learning and contextual memory tasks were unaffected. Definitions of variables may be found below.

The G2CMine data warehouse provides cohort summaries and individual mouse observations from the SAP102 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=11)	Wildtype F (n=0)	Mutant M (n=11)	Mutant F (n=0)	P(sex×mutation)	P(mutation)
EPM total distance	cm	958 (70)	-	723 (99)	-	-	0.066
EPM max speed	cm/s	19 (1.1)	-	17.2 (0.9)	-	-	0.22
EPM % time in open	%	20.5 (6.2)	-	46.7 (11.3)	-	-	0.055
EPM time in centre	s	139 (14)	-	157 (19)	-	-	0.44
EPM max speed, open vs closed	cm/s	-10.3 (1.6)	-	-4 (1.8)	-	-	0.014 *
OF, NOE total distance	log10 cm	3.11 (0.08)	-	2.84 (0.04)	-	-	0.0071 **
NOE vs OF distance travelled	cm	-327 (144)	-	-80 (49)	-	-	0.12
RR naive fall time	log10 s	0.73 (0.12)	-	1.16 (0.08)	-	-	0.0079 **
RR learning	s/trial	1.4 (0.8)	-	1.2 (1.4)	-	-	0.94
RR memory	s	9.1 (5.4)	-	-3.5 (4.2)	-	-	0.079
Fear learning, trial effect	% freezing	53.2 (8.7)	-	42.3 (11.8)	-	-	0.46
Fear learning, tone effect	% freezing	-14.2 (7.1)	-	-22.7 (9.9)	-	-	0.5
Contextual memory, mean	% freezing	51.2 (8.8)	-	43.1 (8.5)	-	-	0.52
Contextual memory, change	% freezing	13.7 (9.2)	-	13.9 (7.9)	-	-	0.99
Cued memory, mean	% freezing	-9.4 (8.1)	-	-0.6 (9.6)	-	-	0.49
Cued memory, change	% freezing	3.1 (6.6)	-	25.3 (6)	-	-	0.021 *

Elevated Plus Maze

 

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=11)	Wildtype F (n=0)	Mutant M (n=11)	Mutant F (n=0)	P(sex×mutation)	P(mutation)
EPM total distance	cm	958 (70)	-	723 (99)	-	-	0.066
EPM max speed	cm/s	19 (1.1)	-	17.2 (0.9)	-	-	0.22
EPM % time in open	%	20.5 (6.2)	-	46.7 (11.3)	-	-	0.055
EPM time in centre	s	139 (14)	-	157 (19)	-	-	0.44
EPM max speed, open vs closed	cm/s	-10.3 (1.6)	-	-4 (1.8)	-	-	0.014 *

Open Field/Novel Object

 

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=11)	Wildtype F (n=0)	Mutant M (n=11)	Mutant F (n=0)	P(sex×mutation)	P(mutation)
OF, NOE total distance	log10 cm	3.11 (0.08)	-	2.84 (0.04)	-	-	0.0071 **
NOE vs OF distance travelled	cm	-327 (144)	-	-80 (49)	-	-	0.12

Rotarod

 

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=11)	Wildtype F (n=0)	Mutant M (n=11)	Mutant F (n=0)	P(sex×mutation)	P(mutation)
RR naive fall time	log10 s	0.73 (0.12)	-	1.16 (0.08)	-	-	0.0079 **
RR learning	s/trial	1.4 (0.8)	-	1.2 (1.4)	-	-	0.94
RR memory	s	9.1 (5.4)	-	-3.5 (4.2)	-	-	0.079

Fear Conditioning

 

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=11)	Wildtype F (n=0)	Mutant M (n=11)	Mutant F (n=0)	P(sex×mutation)	P(mutation)
Fear learning, trial effect	% freezing	53.2 (8.7)	-	42.3 (11.8)	-	-	0.46
Fear learning, tone effect	% freezing	-14.2 (7.1)	-	-22.7 (9.9)	-	-	0.5
Contextual memory, mean	% freezing	51.2 (8.8)	-	43.1 (8.5)	-	-	0.52
Contextual memory, change	% freezing	13.7 (9.2)	-	13.9 (7.9)	-	-	0.99
Cued memory, mean	% freezing	-9.4 (8.1)	-	-0.6 (9.6)	-	-	0.49
Cued memory, change	% freezing	3.1 (6.6)	-	25.3 (6)	-	-	0.021 *