Table 1 Phenotype of Rapgef2-cKO and Rapgef6-KO mice in comprehensive behavioral test battery

Physical characteristics

Body weight

Weight (g)

n.s.

– –

F_1,43 = 1.745, p = 0.1935

F_1,38 = 12.541, p = 0.0011

Fig. 1a, e

Body temperature

Temperature (°C)

n.s.

n.s.

F_1,43 = 1.991, p = 0.1654

F_1,38 = 4.036, p = 0.0517

Fig. 1b, f

Grip strength (N)

Strength (N)

n.s.

n.s.

F_1,43 = 1.149, p = 0.2897

F_1,38 = 0.062, p = 0.8053

Fig. 1c, g

Wire hang latency (s)

Latency (s)

– –

n.s.

F_1,43 = 16.785, p = 0.0002

F_1,38 = 0.04, p = 0.8425

Fig. 1d, h

Motor coordination

Rotarod

Latency to fall (s)

n.s.

n.s.

F_1,43 = 1.251, p = 0.2695

F_1,38 = 0.708, p = 0.4052

Fig. 1i, j

Pain sensitivity

Hot plate

Latency (s)

–

n.s.

F_1,43 = 5.043, p = 0.0299

F_1,38 = 1.045, p = 0.3131

Fig. 1k, l

Exploratory locomotion

Open field test

Distance traveled (cm)

+ + +

+ (initial phase)

F_1,43 = 36.348, p < 0.0001

F_1,38 = 0.03, p = 0.8638

Fig. 2a, c

Anxiety-like behavior

Open field test

Center time (s)

+ + +

+ (initial phase)

F_1,43 = 19.658, p < 0.0001

F_1,38 = 0.002, p = 0.9656

Fig. 2b, d

Light/dark transition test

Number of transitions

n.s.

n.s.

F_1,43 = 0.735, p = 0.3961

F_1,38 = 0.395, p = 0.5335

Fig. 2e, g

Latency to light chamber (s)

n.s.

n.s.

F_1,43 = 0.501, p = 0.4827

F_1,38 = 0.669, p = 0.4187

Fig. 2f, h

Elevated plus maze test

Entries into open arms (%)

+ +

n.s.

F_1,42 = 8.453, p = 0.0058

F_1,38 = 1.328, p = 0.2564

Fig. 2i, k

Time on open arms (%)

+

n.s.

F_1,42 = 6.229, p = 0.0166

F_1,38 = 0.095, p = 0.7597

Fig. 2j, l

Behavioral despair

Porsolt forced swim test

Immobility (%) on Day 1

n.s.

- (1-7 min)

F_1,43 = 2.71, p = 0.107

F_1,38 = 1.881, p = 0.1783;

1-7 min, F_1,38 = 5.089, p = 0.0299

Fig. 3a, b

Immobility (%) on Day 2

- (1-3 min),

+ + (4-10 min)

- (1-2 min)

F_1,43 = 0.065, p = 0.8007;

1-3 min, F_1,43 = 6.82, p = 0.0124;

4-10 min, F_1,43 = 17.252, p = 0.0002

F_1,38 = 2.952, p = 0.0939;

1-2 min, F_1,38 = 4.865, p = 0.0335

Fig. 3a, b

Tail suspension test

Immobility (%)

+ + (7-10 min)

n.s.

F_1,43 = 3.569, p = 0.0656;

7-10 min, F_1,43 = 8.574, p = 0.0054

F_1,38 = 2.651, p = 0.1118

Fig. 3c, d

Sensorimotor gating

Startle response test

Startle response

–

–

F_1,43 = 4.129, p = 0.0484

F_1,38 = 6.287, p = 0.0166

Fig. 4a, c

Prepulse inhibition test

PPI (startle stimulus, 110 dB)

n.s.

n.s.

F_1,43 = 1.161, p = 0.2873

F_1,38 = 1.007, p = 0.322

Fig. 4b, d

PPI (startle stimulus, 120 dB)

n.s.

n.s.

F_1,43 = 0.293, p = 0.5912

F_1,38 = 0.222, p = 0.6404

Fig. 4b, d

Social interaction

Novel environment

Total duration of contacts (s)

n.s.

n.s.

F_1,18 = 0.048, p = 0.8299

F_1,18 = 0.215, p = 0.6484

Fig. 5a, f

Number of contacts

+ + +

+ +

F_1,18 = 28.989, p < 0.0001

F_1,18 = 9.088, p = 0.0074

Fig. 5b, g

Total duration of active contacts (s)

+ + +

n.s.

F_1,18 = 27.519, p < 0.0001

F_1,18 = 4.132, p = 0.0571

Fig. 5c, h

Mean duration of contact (s)

–

–

F_1,18 = 5.566, p = 0.0298

F_1,18 = 5.719, p = 0.0279

Fig. 5d, i

Distance traveled (cm)

+ + +

+

F_1,18 = 39.017, p < 0.0001

F_1,18 = 6.421, p = 0.0208

Fig. 5e, j

Home cage

Mean number of particles

n.s.

n.s.

F_1,16 = 0.743, p = 0.4016;

light period, F_1,16 = 0.031, p = 0.8628;

dark period, F_1,16 = 1.65, p = 0.2173

F_1,18 = 0.002, p = 0.9695;

light period, F_1,18 = 0.438, p = 0.5163;

dark period, F_1,18 = 0.143, p = 0.7097

Fig. 5k, m

Activity levels

n.s.

n.s.

F_1,16 = 0.141, p = 0.712;

light period, F_1,16 = 0.928, p = 0.3497;

dark period, F_1,16 = 0.546, p = 0.4705

F_1,18 = 0.031, p = 0.8633;

light period, F_1,18 = 0.96, p = 0.3401;

dark period, F_1,18 = 0.169, p = 0.6858

Fig. 5l, n

Crawley’s version

Time spent around cage (s) on 1st

– –

n.s.

F_1,43 = 8.754, p = 0.005

F_1,38 = 1.221, p = 0.276

Fig. 5o, p

Time spent around cage (s) on 2nd

– –

n.s.

F_1,43 = 9.539, p = 0.0035

F_1,38 = 0.014, p = 0.9074

Fig. 5o, p

Spatial memory

Barnes maze test

Acquisition test

Latency to 1st (s)

n.s.

n.s.

F_1,43 = 2.556, p = 0.1172

F_1,38 = 0.0004, p = 0.9841

Fig. 6a, n

Error to 1st

+ + +

n.s.

F_1,43 = 18.985, p < 0.0001

F_1,38 = 0.657, p = 0.4228

Fig. 6b, o

Distance to 1st (mm)

+ + +

n.s.

F_1,43 = 18.746, p < 0.0001

F_1,38 = 0.668, p = 0.4187

Fig. 6c, p

Probe test (1-day after training)

Latency to 1st (s)

+ +

n.s.

F_1,43 = 8.983, p = 0.0045

F_1,38 = 1.506, p = 0.2273

Fig. 6d, q

Error to 1st

+ +

n.s.

F_1,43 = 13.605, p = 0.0006

F_1,38 = 0.463, p = 0.5005

Fig. 6e, r

Distance to 1st (mm)

+ +

n.s.

F_1,43 = 16.909, p = 0.0002

F_1,38 = 0.544, p = 0.4653

Fig. 6f, s

Time spent around each hole (s)

n.s.

n.s.

F_1,43 = 0.062, p = 0.8026

F_1,38 = 0.35, p = 0.5576

Fig. 6g, t

Time spent around target hole (s)

– – –

n.s.

F_1,43 = 23.594, p < 0.0001

F_1,38 = 0.3912, p = 0.0552

Fig. 6h, u

Probe test (1-month after training)

Latency to 1st (s)

n.s.

n.s.

F_1,42 = 0.488, p = 0.4885

F_1,38 = 1.705, p = 0.1995

Fig. 6i, v

Error to 1st

+ +

n.s.

F_1,42 = 9.308, p = 0.0039

F_1,38 = 2.786, p = 0.1033

Fig. 6j, w

Distance to 1st (mm)

+

n.s.

F_1,42 = 5.991, p = 0.0186

F_1,38 = 4.029, p = 0.0519

Fig. 6k, x

Time spent around each hole (s)

n.s.

n.s.

F_1,42 = 6.468, p = 0.0148

F_1,38 = 0.021, p = 0.886

Fig. 6l, y

Time spent around target hole (s)

n.s.

n.s.

F_1,43 = 0.317, p = 0.5762

F_1,38 = 1.853, p = 0.1814

Fig. 6m, z

Working memory

T-maze test

Correct responses (%)

– – –

- - (delayed alternation)

F_1,43 = 30.696, p < 0.0001

Session 1-4, F_1,38 = 1.361, p = 0.2506

Session 5-8, F_1,38 = 7.996, p = 0.0074

Fig. 7a, c

Cued and contextual fear conditioning

Fear conditioning test

Conditioning

Freezing (%)

– – –

– –

F_1,43 = 32.949, p < 0.0001

F_1,38 = 10.046, p = 0.003

Fig. 8a, g

Context test 1 day after conditioning

Freezing (%)

– – –

–

F_1,43 = 36.049, p < 0.0001

F_1,38 = 6.837, p = 0.0127

Fig. 8b, h

Cued test 1 day after conditioning (pre-CS)

Freezing (%)

–

– –

F_1,43 = 4.641, p = 0.0369

F_1,38 = 10.947, p = 0.0021

Fig. 8b, h

Cued test 1 day after conditioning (CS)

Freezing (%)

n.s.

–

F_1,43 = 4.003, p = 0.0517

F_1,38 = 5.287, p = 0.0271

Fig. 8b, h

Context test 1 month after conditioning

Freezing (%)

– – –

–

F_1,42 = 36.559, p < 0.0001

F_1,38 = 4.772, p = 0.0352

Fig. 8c, i

Cued test 1 month after conditioning (pre-CS)

Freezing (%)

–

– –

F_1,41 = 6.521, p = 0.0145

F_1,38 = 15.314, p = 0.0004

Fig. 8c, i

Cued test 1 month after conditioning (CS)

Freezing (%)

–

–

F_1,41 = 5.188, p = 0.028

F_1,38 = 5.254, p = 0.0275

Fig. 8c, i

Fear conditioning test (foot shock 1)

Distance traveled (cm)

n.s.

+ +

F_1,43 = 10.489, p = 0.4883

F_1,38 = 12.428, p = 0.0011

Fig. 8d, j

Fear conditioning test (foot shock 2)

Distance traveled (cm)

n.s.

n.s.

F_1,43 = 0.423, p = 0.5189

F_1,38 = 0.347, p = 0.5592

Fig. 8d, j

Fear conditioning test (foot shock 3)

Distance traveled (cm)

n.s.

n.s.

F_1,43 = 1.026, p = 0.3168

F_1,38 = 1.858, p = 0.1809

Fig. 8d, j

Context test 1 day after conditioning

Activity suppression ratio

+ +

n.s.

F_1,43 = 15.048, p = 0.0004

F_1,38 = 1.699, p = 0.2002

Fig. 8e, k

Context test 1 month after conditioning

Activity suppression ratio

+ + +

n.s.

F_1,42 = 46.445, p < 0.0001

F_1,38 = 0.154, p = 0.697

Fig. 8e, k

Cued test 1 day after conditioning (CS)

Activity suppression ratio

n.s.

n.s.

F_1,43 = 0.211, p = 0.6482

F_1,38 = 1.274, p = 0.2662

Fig. 8f, l

Cued test 1 month after conditioning (CS)

Activity suppression ratio

n.s.

n.s.

F_1,41 = 3.032, p = 0.0891

F_1,38 = 1.046, p = 0.3129

Fig. 8f, l