Inducing Depression-like Behavior in a Mouse via Immobilization Stress

Protokol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Induction of depression by CIS restraint

NOTE: Handle the mouse gently but firmly with confidence. Rough and tentative handling is another stress factor in the experiment, and it is important for the mouse's struggling, biting, and scratching.

1. Set the room light to light (200 Lux) conditions using a digital lux meter.
2. House the mouse in a separate cage at least a week before testing and place the mouse in the testing room for at least 30 min before the experiment.
   NOTE: Handle the mice at least once a day for at least three consecutive days before the experiment so that they become familiar with the experimenter. An adaptational period before the experiment is necessary to ensure that the mice acclimatize to the circumstance, such as the testing room.
3. Gently hold the mouse tail to avoid tensing the mouse, and then carefully place it on a rough surface (top of the wire bar of the cage or cage lid).
4. Cover the restrainer with a small white towel, and then gently place the mouse at its opening so that it enters the restrainer voluntarily.
   NOTE: In this case, the mouse is positioned in the opposite direction from where it enters the restrainer. To encourage the mouse to enter the restrainer voluntarily, the restrainer is covered with a small towel to make the inside darker.
5. Place the closure to restrain the mouse as tight as possible, being careful to avoid damage to the body, such as the tail, feet, and testicles.
6. Restrain the mouse for 2 h/day (9:00 A.M. to 11:00 A.M.) for 15 consecutive days.

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Malzemeler

Name	Company	Catalog Number	Comments
Ball nozzle	Jeung Do B&P	JD-C-88
CCTV camera	KOCOM	KCB-381
Digital lux meter	TES	TES-1330A
Mice	Koatech		C57BL/6 strain
Restrainer	Dae-jong Instrument Industry	DJ-428
Water bottle	Jeung Do B&P	JD-C-79