To design a Standard Operating Procedure that describes the procedure for operation and maintenance of the Elevated Plus Maze.
2.0 SCOPE
This Standard Operating Procedure (SOP) is applicable for the operation, and maintenance of elevated plus maze Pharmacology studies conducted at animal house facility
3.0 RESPONSIBILITY
3.1 All personnel in the Pharmacology Department shall be responsible for implementation and compliance with the SOP.
3.2 Head/Designees/Scientists/Study Directors of the Pharmacology department shall be responsible to ensure the implementation and compliance with the SOP.
4.0 DISTRIBUTION
4.1 The quality assurance department is responsible to keep SOP ‘Master Copy’ approved through the Quality Assurance Head.
4.2 The copy of ‘Control Copy’ of all SOPs is being distributed in the pharmacology department and placed near related Equipment/Instruments as a ‘Display Copy’.
5.0 DEFINITION(S)
5.1 Definition(s)
5.1.1 The elevated plus maze has been described as a simple method for assessing the anxiety responses of rodents.
5.1.2 The elevated plus maze has predictive validity, which is defined as the extent to which the dependent measure predicts behavior on a related measure.
6.1 Abbreviation(s)
6.1.1 SOP: Standard Operating Procedure
6.1.2 PPEs: Personal Protective Equipment
7.0 PROCEDURE
7.1 Introduction
7.2 Instrument description
7.3 Operations
7.3.5 Behaviors scored included:
7.4.1 Clean the testing arena with soap and water. Allow the arena to dry before use.
7.4.2 Begin recording. Hold the test card (with the date, test name, and testing conditions) in view. Hold the subject’s cage card in view.
7.4.3 Place the subject in the center of the arena.
7.4.4 After five minutes, return the subject to its cage.
8.0 PRECAUTIONS
8.1 All PPEs like aprons, gloves, masks, head caps, and shoe covers should be worn before using the instrument.
8.2 Clean the equipment with a dry piece of robe or a lightly moistened one, do not use organic solutions.
8.3 Never use abrasive products or dissolvents.
8.4 Never pour water or liquids on the equipment.
9.0 REFERENCES
9.1 References
9.1.1 Bindra, D., & Thompson, WR. 1953. An evaluation of defecation and urination as measures of fearfulness. Journal of Comparative and Physiological Psychology, 46, 43- 45.
9.1.2 Blanchard, DC, Griebel, G., Blanchard, RJ. 2001. Mouse defensive behaviors: Pharmacological and behavioral assays for anxiety and panic. Neuroscience and Biobehavioral Reviews, 25, 205-218.
END OF THE DOCUMENT
6.1.1 SOP: Standard Operating Procedure
6.1.2 PPEs: Personal Protective Equipment
7.0 PROCEDURE
7.1 Introduction
7.1.1 This is a commonly used paradigm to test primarily for anxiety-like behavior and locomotor activity in rodents.
7.1.2 The elevated plus maze is a widely used behavioral assay for rodents and it has been validated to assess the anti-anxiety effects of pharmacological agents and steroid hormones, and to define brain regions and mechanisms underlying anxiety-related behavior.
7.2.1 The Elevated Plus-Maze was built according to the description of Lister (1987). It had a black Plexiglas floor with a 5 x 5 cm central square platform, from which radiated two 45 x 5 cm open arms with 0.25 cm high edges and two 45 x 5 cm closed arms with 40 cm high walls made of clear Plexiglas. A white line was drawn halfway along each of the four arms and was used to measure locomotion. The apparatus was raised to 45 cm above the floor on a plus-shaped plywood base. The apparatus was located in a 2 x 5 m laboratory room that was illuminated with a 60-watt red light bulb.
7.3.1 Mice were carried into the test room in their home cages and were handled by the base of their tails at all times. Mice were placed in the central square of the Plus-Maze facing an open arm and were then allowed to explore the apparatus for 5 minutes.
7.3.2 An observer sitting quietly about 1 m from the apparatus recorded the behavior of the animals in the maze. A video camcorder located 150 cm above the center of the maze also recorded behavior.
7.3.3 Rearing, head-dips, grooming, stretch attends and line crosses were scored by the experimenter, whereas measures of arm entries and the time in each arm were recorded with the Limelight video camera-based tracking system (Actimetrics), which used a video camera positioned 2.1 m above the apparatus.
7.3.4 After 5 minutes, mice were removed from the maze by the base of their tails and returned to their home cage. The maze was then cleaned with a solution of 70% ethyl alcohol and permitted to dry between tests.
7.3.6 Open arms entries: Frequency with which the animal entered the open arms. All four of the mouse’s paws were required to be in the arm to be counted as an entry.
7.3.7 Closed arm entries: Frequency with which the animal entered the closed arms. All four of the mouse’s paws were required to be in the arm to be counted as an entry
7.3.8 Open arm duration: Length of time the animal spent in the open arms.
7.3.9 Closed arm duration: Length of time the animal spent in the closed arms.
7.3.10 Center square entries: Frequency with which the animal entered the central square with all four paws.
7.3.11 Central square duration: Length of time the animal spent in the central square.
7.3.12 Head dipping: Frequency with which the animal lowered its head over the sides of the open arm toward the floor.
7.3.13 Stretch attends postures: Frequency with which the animal demonstrates forward elongation of head and shoulders followed by retraction to the original position.
7.3.14 Rearing: Frequency with which the animal stands on its hind legs or leans against walls of the maze with front paws.
7.3.15 Grooming: Frequency and duration of time the animal spent licking or scratching itself while stationary.
7.3.16 Urination: Number of puddles or streaks of urine.
7.3.17 Defecation: Number of fecal boli produced.
7.3.18 Locomotion: Number of times the animal crossed a line drawn on the open and closed arms.
7.3.19 The percentage of entries into the open arms and closed arms based on the total arms entries was also calculated for each animal. The percentage of time spent in the open arms and the closed arms was calculated over the 5-minute test.
7.4 Maintenance
7.4.1 Clean the testing arena with soap and water. Allow the arena to dry before use.
7.4.2 Begin recording. Hold the test card (with the date, test name, and testing conditions) in view. Hold the subject’s cage card in view.
7.4.3 Place the subject in the center of the arena.
7.4.4 After five minutes, return the subject to its cage.
8.0 PRECAUTIONS
8.1 All PPEs like aprons, gloves, masks, head caps, and shoe covers should be worn before using the instrument.
8.2 Clean the equipment with a dry piece of robe or a lightly moistened one, do not use organic solutions.
8.3 Never use abrasive products or dissolvents.
8.4 Never pour water or liquids on the equipment.
9.0 REFERENCES
9.1 References
9.1.1 Bindra, D., & Thompson, WR. 1953. An evaluation of defecation and urination as measures of fearfulness. Journal of Comparative and Physiological Psychology, 46, 43- 45.
9.1.2 Blanchard, DC, Griebel, G., Blanchard, RJ. 2001. Mouse defensive behaviors: Pharmacological and behavioral assays for anxiety and panic. Neuroscience and Biobehavioral Reviews, 25, 205-218.
END OF THE DOCUMENT
You may like to read these links:
1. List of All SOPs and Documents for the Microbiology Laboratory
2. List of All SOPs and Documents for In-vitro Laboratory
3. List of All SOPs and Documents for the Animal House Facility
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