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Place an isolated and acclimatized mouse in a testing room under moderate lighting to avoid sudden stress.
In a stress-free state, the brain neurons release glutamate, an excitatory neurotransmitter that binds to the receptors on neighboring neurons, facilitating signal transmission.
Gently hold the mouse by the tail to prevent tensing or muscle tightening, then place it on a rough surface.
Next, position the mouse near a covered restrainer, encouraging its voluntary entry into the dark hole within the restrainer.
Immobilize the mouse in the restrainer without injury, inducing immobilization stress.
Repeat this immobilization process over multiple sessions to induce chronic stress, triggering the release of corticosterone, a stress hormone.
Corticosterone diffuses inside the neurons and binds to their glucocorticoid receptors, triggering intracellular signaling that reduces the number of glutamate receptors.
This reduces glutamate response and signal transmission, leading the mouse to develop depression-like behaviors with a decreased interest in activities.
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