PHARMACOLOGY 2 LABORATORY

DATE:

AGUSTIN, ROSSET FAYE

BELIGAN, AIRISH
LAGMAY, EFRELYN MAE
TADENA, MARIMAR

CNS Stimulation on Laboratory Mice

I. CNS Stimulation
A. The Open Field

Test Procedure:

Preparation: The Open Field Test requires a square arena with high walls to prevent mice from
escaping. The floor of the arena is divided into a grid of squares to facilitate recording of the mouse's
movements. Before the experiment, the arena is cleaned and dried to remove any potential odors or
stimuli that may affect the mouse's behavior.

Acclimation: Before the actual test, the mice are placed in the testing room for at least 30 minutes to
allow them to acclimate to the environment.

Testing: The mouse is placed in the center of the arena and its behavior is observed for a predetermined
period of time, typically ranging from 5 to 30 minutes. The researcher records the time spent in the center
of the arena (anxiety-like behavior) versus the time spent in the periphery (exploratory behavior) as well
as the distance traveled by the mouse.

B. The Elevated Plus Maze

Test Procedure:

Preparation: The Elevated Plus Maze is a plus-shaped platform with two open arms and two enclosed
arms. The arms are elevated above the ground, creating a potential threat for the mouse and causing
anxiety-like behavior. The maze is located in a quiet and well-lit room to reduce the influence of
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environmental factors on the mouse's behavior.

Acclimation: Before the actual test, the mice are placed in the testing room for at least 30 minutes to
allow them to acclimate to the environment.

Testing: The mouse is placed in the center of the maze facing one of the enclosed arms, and its behavior
is observed for a predetermined period of time, typically ranging from 5 to 10 minutes. The researcher
records the time spent in the open arms (exploratory behavior) versus the time spent in the enclosed arms
(anxiety-like behavior) as well as the number of entries into each arm.
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C. The Novel Object Test

Preparation: The test requires a testing arena, such as an open-field box or a T-maze, and two objects
of similar size and shape. The objects should be novel to the mouse and should not have been used in
prior tests. The arena is located in a quiet and well-lit room to reduce the influence of environmental
factors on the mouse's behavior.

Acclimation: Before the actual test, the mice are placed in the testing room for at least 30 minutes to
allow them to acclimate to the environment.

Training: The mice are placed in the testing arena with two identical objects and allowed to explore
them for a predetermined period of time, typically ranging from 5 to 10 minutes. This is known as the
familiarization phase.

Delay: After the familiarization phase, the mice are removed from the arena and a delay period is
introduced, typically ranging from 1 to 24 hours. This delay period allows the mouse to forget familiar
objects.

Testing: The mouse is placed back in the arena with one familiar object and one novel object. The time
spent exploring each object is recorded, typically ranging from 5 to 10 minutes. The researcher records
the amount of time spent exploring the novel object versus the familiar object.
PHARMACOLOGY 2 LABORATORY

DATE:

AGUSTIN, ROSSET FAYE

BELIGAN, AIRISH
LAGMAY, EFRELYN MAE
TADENA, MARIMAR

1. Worksheet for CNS Stimulation

Name of the Test

Mouse 1 observation Mouse 2 observation
A. The Open Field Test

Time spent in the center of

3 seconds 1 second
the arena

Total time spent at

the periphery 2 seconds 1 second

23 x 32 cm 23 x 32 cm
Maximum Distance traveled

B. The Elevated Plus

Maze

Exploratory Behavior
in Minutes 6 seconds 49 seconds

Anxiety-like behavior
4 minutes and 54 seconds 4 minutes and 11 seconds
in minutes

Number of entries the

mice stayed on the open 1 entry 3 entries
arms of the maze

Number of entries the mice

stayed on the closed arms of 5 entries 5 entries
the maze

C. The Novel Object

Test
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The time when the

mice explore the 5 seconds 1 second
novel object
The time when the mice
explore the familiar object 4 seconds 2 seconds

DOCUMENTATION

● Measuring the box

Size: 23 x 32 cm

A. The Open Field Test

Mouse 1 observation
ACCLIMATION TESTING
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Mouse 2 observation
ACCLIMATION TESTING

B. The Elevated Plus Maze

Mouse 1 observation
ACCLIMATION
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TESTING

Open arms (exploratory behavior) Enclosed arms (anxiety-like behavior)

Mouse 2 observation
ACCLIMATION

TESTING
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Open arms (exploratory behavior) Enclosed arms (anxiety-like behavior)

C. The Novel Object Test

Mouse 1 observation
ACCLIMATION TRAINING DELAY

TESTING TESTING
Familiar Object: Binder Clip Novel Object: Eraser

Mouse 2 observation
ACCLIMATION TRAINING DELAY
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TESTING TESTING
Familiar Object: Pen Cap Novel Object: Eraser