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Protocol     ・List of test items      (Ver. 4, 18KB) No. In the Viewing Jar Observation in the viewing jar. Ver. 1 Ver. 2 Ver. 3 Ver. 4 Ver. 5 Q1 Coat color Determine the hair color of the mouse. Q1.1 Major color Determine the hair color of the large part of the mouse's body. Q1.2 Minor color Determine the hair color of the small part of the mouse's body. Q1.4 Pattern in the region with a different color Determine the pattern in the region with a different color. Q1.5 White belly Determine the kind of white belly. Q2 Hair length Examine hair length. Q3 Hair morphology Examine the morphological characteristics of the mouse's body hair. Q63 Injury Determine skin injury. Q4 Respiration rate Determine the respiration rate after the five minutes. Q5 Tremor Determine whether the mouse shakes. Q6 Body position Observe the posture and position of the mouse. Q7 Spontaneous activity Observe the degree of activity of the mouse's movement. Q8 Defecation Leave the mouse for five minutes, and record the number of feces. Q9 Urination Leave the mouse for five minutes, and see if the mouse has urinated. In the Arena Observation in the arena. Q10 Time elapsed before the mouse starts to move (seconds) Make a note of the time elapsed before the mouse starts to move. Q11 Locomotor activity Make a note of the total number of squares entered in first 30 seconds. Q12 Transfer arousal Evaluate the behavior of the mouse in the arena in first 30 seconds. Q13 Piloerection Observe body hair and piloerection. Q15 Startle response Evaluate how the mouse responds to the auditory stimulus of the IHR click box. Q16 Gait Observe the gait of the mouse. Q17 Pelvic elevation Observe the height of the pelvis from the arena floor while walking. Q18 Tail elevation Determine how the position of the tail. Q19 Touch escape Approach the mouse with a finger and observe the response. Above the Arena 1 Observation above the arena. Q20 Positional passivity Hold the mouse by the several ways on the arena and observe whether it resists or not. Q21 Trunk curl Determine whether the mouse curls its body forward when dangled by the tail. Q22 Limb grasping Determine whether the mouse holds its forelimbs and hindlimbs together or either of them when dangled by the tail. On the Arena Observation on the grid (on the arena). Q23 Visual placing Hold the mouse by the tail and lower it onto the grid. Determine the distance between the mouse and the grid when it extends its forelimbs to the grid. Q24 Grip strength Hold the mouse by the tail and drag it on the grid. Evaluate the strength of the mouse’s grip. Q25 Body tone Compress both sides of the mouse between the thumb and forefinger. Evaluate the degree of resistance. Q26 Head morphology Observe the morphology of the mouse's head. Q27 Pinna reflex Stimulate the base of the pinna with the wire probe. Record how the pinna moves. Q28 Pinna morphology ( R ) Examine the morphology of the right pinna. Q29 Pinna morphology ( L ) Examine the morphology of the left pinna. Q30 Corneal reflex Stimulate the cornea softly with the curved part of the wire and observe whether the mouse blinks. Q31 Toe pinch Use curved forceps to squeeze one finger of the hindlimb. Observe the response. Above the Arena 2 Observation while holding the mouse. Q32 Body length Measure the distance from the tip of the nose to the tail base with a ruler. Q33 Tail length Measure the distance from the tail base to the tip of the tail with the ruler. Q34 Tail morphology Examine the morphology of the tail. Q35 Lacrimation Restrain the mouse and determine whether tears are secreted. Q14 Palpebral closure Examine both eyes and their degree of closure. Q62 Palpebral fissure length Observe the length of the palpebral fissures. Q59.1 Cornea Observe the cornea of the mouse. Q59.2 Pupil Observe the pupil of the mouse. Q59.3 Eye size Observe the eye size of the mouse. Q59.4 Eye color Observe the eye color of the mouse. Q59.5 Eye shape Observe the shape of eye of the mouse. Q36 Whisker morphology Examine the morphology of the whiskers (sensory hairs). Q37 Tooth morphology Observer and examine the morphology of the incisor. Q38 Provoked biting Approach the mouth with a plastic rod and observe whether the mouse bites the rod. Q39 Salivation Examine the plastic rod used in Q38 to determine how much saliva adhered to the rod. Q40 Heart rate Place a forefinger on the chest of the mouse and feel the heartbeat. Q41 Abdominal tone Gently press on the abdomen of the mouse with a forefinger and evaluate the degree of resistance. Q42 Skin color Examine the color of the ventral aspects of the four footpads. Q43-46 Limb morphology (FR, FL, HR, HL) Examine the shape of four limbs. Q47 Limb tone Press against the sole of the mouse's hindlimbs with a forefinger and middle finger and evaluate how strong the mouse makes resistance. Q48 Wire maneuver Release the mouse on the wire grasping with only its forelimbs and observe whether the mouse can grasp the wire with its hindlimb. Q49 Righting reflex Observe the righting reflex. Q50 Contact righting reflex Observe the righting reflex in the tube. Q51 Negative geotaxis Put the mouse on the grid and turn the grid to let it to be oriented with its head down. Observe how the mouse moves in response to the orientation. Additional Comments and Body Ratio Parameters Observation of whole test and calculation of body ratio parameters. Q52 Fear Determine if the mouse freezes whenever it is transferred from one place to another. Q53 Irritability Determine whether the mouse violently resists being restrained during the tests. Q54 Aggression Determine whether the mouse defied and bit the handler during the tests. Q55 Vocalization Determine whether the mouse squeaked during the tests. Q56 Bizarre behaviour Determine whether the mouse shows any bizarre behaviors. Q57 Convulsions Determine whether the mouse exhibits any convulsions. Q58 Body weight Weigh the mouse on a scale. Q60 Body mass index (BMI) Calculated BMI from body length and body weight. Q61 Tail ratio (Tr) Calculated Tr from body length and tail length.

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