Procedures

Procedure 1: Fluoxetine administration

Reagents and solutions

• Fluoxetine HCl (Medisca Inc., Lot 172601, Plattsburgh, NY, USA)

Environmental Conditions

Holding Area and/or Sample Collection Area

Acoustic Background: Ambient

Steps

1. The average weight measurements for each strain, together with previously determined daily water intake for each strain, are used to determine the amount of fluoxetine required to provide a daily oral dose of 0 (control) or 18 mg/kg/day per mouse in drinking water.
2. Mice are treated daily throughout the experiment, starting at 5-6 wks of age. Content of water bottles is changed every week.

Additional ARRIVE Guidelines

Sample Size Estimate: 10 per treatment group

Allocation of Animals to Test Group: Randomized

Procedure 2: Body weight

Equipment, software, and supplies

• Scales

Environmental Conditions

Holding Area and/or Sample Collection Area

Facility: The Jackson Laboratory's Center for Biometric Analysis (CBA) animal room

Max # Animals Per Enclosure: 5

Acoustic Background: Ambient

Collection Method: Body weight

Additional ARRIVE Guidelines

Sample Size Estimate: 10 mice per treatment group

Allocation of Animals to Test Group: Randomized

Procedure 3: Grip strength test

Equipment, software, and supplies

• Mesh grid
• Grip strength meter (Bioseb, Pinellas Park, FL, USA)

Reagents and solutions

• 70% ethanol, for cleaning apparatus between subjects

Environmental Conditions

Testing Area

Facility: The Jackson Laboratory's Center for Biometric Analysis (CBA) testing room

Acclimation Period: at least 60 min

Acoustic Background: Ambient

Collection Method: Whole Mouse

Female and Male Mice Tested in Same Test Group: No

Sanitation of Equipment: 70% ethanol

Steps

1. Mice are acclimated for 60 min prior to testing.
2. Mice are weighed prior to testing to adjust measures for body weight where noted.
3. Each mouse is held by the tail, lowered over the grid -- keeping the torso horizontal and allowing either only its forepaws to attach or all four limbs to attach -- before any measurements are taken.
4. Each mouse is gently pulled back by its tail, ensuring the grip on the top portion of the grid, with the torso remaining horizontal.
5. A grip strength meter measures the grip.

Additional ARRIVE Guidelines

Sample Size Estimate: 10 mice per treatment group

Allocation of Animals to Test Group: Randomized

Procedure 4: Tail suspension test

Equipment, software, and supplies

• Tail suspension test chambers (Med Associates, St. Albans, VT, USA)
• Noldus Ethovision (XT15)
• Adhesive tape, 25 mm wide (Fisher)

Reagents and solutions

• 70% ethanol, for cleaning apparatus between subjects

Environmental Conditions

Testing Area

Facility: The Jackson Laboratory's Center for Biometric Analysis (CBA) testing room

Acclimation Period: at least 60 min

Acoustic Background: Ambient

Collection Method: Whole Mouse

Female and Male Mice Tested in Same Test Group: No

Sanitation of Equipment: 70% ethanol

Steps

1. Mice are allowed to acclimate to the testing room for a minimum of 60 min.
2. Each mouse is suspended by its tail with adhesive tape to a flat metal bar connected to a force transducer that measures transduced movement. The tape is extended the length of the tail from 2 mm from the base through 2 mm from the tip, minimizing the ability of the mouse to climb its tail. Mice that climb their tails are excluded.
3. A computer interfaces with the force transducer to record the data. Highly mobile: above 60%, immobile: below 9%, mobile: in between highly mobile and immobile. If any part of the subject is occupying a cell contained by a zone, the subject is considered to be ‘inside’ that zone.

Additional ARRIVE Guidelines

Sample Size Estimate: 10 mice per treatment group

Allocation of Animals to Test Group: Randomized

Data Inclusion & Exclusion Criteria: Mice excluded if they climbed their tails

Procedure 5: Open field test

Equipment, software, and supplies

• Open field test apparatus (OmniTech Electronics, Inc., Columbus, OH, USA)

Reagents and solutions

• 70% ethanol, for cleaning apparatus between subjects

Environmental Conditions

Testing Area

Facility: The Jackson Laboratory's Center for Biometric Analysis (CBA) testing room

Acclimation Period: at least 60 min

Acoustic Background: Ambient

Collection Method: Whole Mouse

Female and Male Mice Tested in Same Test Group: No

Sanitation of Equipment: 70% ethanol

Steps

1. Mice are allowed to acclimate to the testing room for a minimum of 60 min.
2. Each mouse is placed in the center of the arena at the start of the test.
3. Data are recorded via sensitive photobeams and collected in 5-min bins.

Additional ARRIVE Guidelines

Sample Size Estimate: 10 mice per treatment group

Allocation of Animals to Test Group: Randomized