Transcript Basic Mouse Neurobehavioral Exam (or giving a mouse a physical)

Basic Mouse Neurobehavioral
Exam
(or how to give a mouse a physical)
Julie Watson Vet MB DACLAM
Resources
• Jacqueline Crawley
– Crawley,J and Paylor,R. Hormones & Behavior 197-211 (1997)
– “What’s Wrong with my Mouse?” Wiley-Liss 2000.
• Samuel Irwin - First Phenotyping Screen
– Psychopharmacologia (Berl.) 13 222-257 (1968)
• SHIRPA stage I
– Mamm Genome. 1997 Oct;8(10):711Rogers DC, Fisher EM, Brown SD, Peters J, Hunter AJ, Martin JE
• Websites:
– Eumorphia
http://www.eumorphia.org/EMPReSS/servlet/EMPReSS.Frameset
– MRC Mutagenesis Program
– http://www.mgu.har.mrc.ac.uk/facilities/mutagenesis/mutabase/
• Clinical approach
Goals
1. Detect abnormalities likely to affect
future behavioral tests
– Blindness
– Physical defects
– Deafness
Background Strain
Age-related hearing
loss & vestibular
defects (<3m)
•
•
•
•
•
Many 129 strains
A/J
C57BR, C57L
DBA
I, LP, NOD, ALR, ALS
• C57BL/6J @12-18m
Blindness
(rd1 gene)
•
•
•
•
•
•
•
•
FVB
C3H
BUB
CBA
SJL
SWR
NON
P, PL
Goals (2)
2. Suggest further testing
– Presence of abnormal behaviors
– Absence of normal behaviors
– Motor or neurological deficits
Pitfalls
• Variations due to:
–
–
–
–
Husbandry methods (see John Crabbe, Science 1999)
Age/sex/background strain
Noise/temperature/humidity/location etc.
Time of day
• Detect only sizeable deficits
– Background strain not inbred. F1’s, N2’s
– Are controls comparable ?
– Insufficient group size
• Limited statistical analysis available - ordinal data
• Check genotyping reliability
Behavioral Phenotyping Level 1 Screen
Accession #_________
Date ________Investigator ____________________Genotype ___________________________________
Background strain(s)_____________Inbred / N#___ Tg /TM KO/KI/Cond
Gene Name_____________
Key: 0 = zero; 1 = slow or reduced; 2 = normal; 3 = hyper
Animal #
WT
Hemi
-/Animal #
WT
DOB/Age
Sex
DOB/Age
Sex
Weight (g)
Condition Score
1..2..3..4
Empty Cage 2 mins:
Gait abnormal
Y
Posture abnormal Y
Freezing
Y
Wild running
Y
Stereotypies
Y
Escape
Y
Fur color
Weight (g)
Condition Score
1..2..3..4
Empty Cage 2 mins:
Gait abnormal
Y N
Posture abnormal Y N
Freezing
Y N
Wild running
Y N
Stereotypies
Y N
Escape
Y N
Fur color
M
F
Exploring
0..1..2..3
N 0= <1 side; 1 =< 1 circuit;
2= multiple circuits; 3= frantic
N
N
Digging
0..1..2..3
N
Grooming
0..1..2..3
N
Rearing
0..1..2..3
N
DORSAL
VENTRAL
_________________
Grooming
0… 1-2
3-6…>6
Digging
0 …1-2
3-6…>6
DRAW
Escape
0…patches/abnormalities
1-2
Bald
3-6
Bald >6
patches?
Y.. N Piloerection?
Physical abnormality Y..N
Whisker damage Y..N
-/M F
Exploring
0..1..2..3
0= <1 side; 1 =< 1 circuit;
2= multiple circuits; 3= frantic
Digging
Grooming
Rearing
DORSAL
Y..N
Hemi
0..1..2..3
0..1..2..3
0..1..2..3
VENTRAL
DRAW
Bald patches/abnormalities
Bald patches?
Y.. N Piloerection?
Physical abnormality Y..N
Y..N
Whisker damage Y..N
Body tone
0..1..2..3 Whisker response
Body tone
0..1..2..3 Whisker response
NA 0..1..2..3
NA 0..1..2..3
Petting escape 0..1..2..3 Ear twitch
0..1..2..3 Petting escape 0..1..2..3 Ear twitch
0..1..2..3
Passivity
0..1..2..3
Trunk curl
0..1..2..3 Forelimb place
Righting
0..1..2..3
Visual placing 0..1..2..3
Palpebral reflex 0..1..2..3
0 ..1...2...3
RL withdraw 0..1..2..3
Biting
Reach c touch 0..1..2..3 Clicker
NA
Grip: >60
Passivity
0..1..2..3 Palpebral reflex 0..1..2..3
Trunk curl
0..1..2..3 Forelimb place 0 ..1..2..3
Righting
0..1..2..3 RL withdraw 0..1..2..3
0..1..2..3 Visual placing 0..1..2..3
Biting
0..1..2..3
0..1..2..3 Reach c touch 0..1..2..3
Clicker
0..1..2..3
<60 time
_______
Grip: >60
<60 time
_______
Notes: _______________________________________________________________________________
____________________________________________________________________________________
BIOSAFETY CABINET
SIMPLE EQUIPMENT
WEIGHT
PERTINENT INFORMATION
TESTER BLINDED TO GENOTYPE
Tests
• Behavior in Empty Cage:
– Gait, posture, general appearance
– Are normal behaviors present?
• Exploring, thigmotaxis, digging, grooming,
rearing
– Are abnormal behaviors present?
• freezing, wild running, stereotypies, seizures,
pruritus
REARING/ESCAPE
DIGGING
Knockout Mouse
LIMITED REARING
Tests
• Pick up, record abnormal physical
features
– Whisker loss
– Bald patches or unkempt haircoat,
piloerection
– Eyes
– Legs and tail
NORMAL WHISKERS
WHISKER LOSS
General Reactivity
•
•
•
•
Response to approach
Body tone
Petting escape
Passivity
Normal Response to Approach
NORMAL BODY TONE
HIGH BODY TONE
PETTING ESCAPE
Unusual Escape Attempts
PASSIVITY
Postural Reactions and Reflexes
•
•
•
•
Trunk curl
Righting reflex
Forelimb proprioceptive positioning
Rearlimb withdrawal
TRUNK CURL
RIGHTING REFLEX
PROPRIOCEPTIVE POSITIONING
WITHDRAWAL - FAST
WITHDRAWAL - SLOW
Tests –V & VII Nerve
Facial sensory and motor
• Whisker response
• Ear twitch
• Palpebral reflex (V, VII)
EAR TWITCH
EAR TWITCH – WHOLE HEAD RESPONSE
WHISKER RESPONSE (?)
PALPEBRAL RESPONSE
Sight
• Placing – visual II
• Placing – tactile
VISUAL PLACING
Tactile Placing
Tests
• Clicker / hearing
• Pupil / response to light
• Biting / aggression
CLICKER
Provoked Aggression
Grip Strength Normal
Grip Strength Abnormal
Impact of Physical Deficits on
Behavioral Tests
Impact of Blindness
• Learning and memory tests
• Water maze, contextual fear conditioning
–
–
–
–
animal cannot learn?
has limited spatial abilities?
Consider alternatives
2 lever or hole-poke test in operant chamber
• Anxiety tests
• Visual cliff, elevated T maze, light- dark box
– Low anxiety?
Whisker Deficits, Itchy Skin
• Lack of Whiskers
– Anxiety tests e.g. visual cliff, elevated T maze –
• Skew response compared with mice with whiskers
• ? Remove whiskers
• Itchy skin (B6 dermatitis)
– open field, aggression
• hyperirritability,
• increased activity
• increased aggression
Hearing Deficits
• Cued fear conditioning
– Use contextual fear conditioning
• Prepulse inhibition
– Nb DBA still works
Examples of Further Testing
•
•
•
•
•
•
Open Field
Light Dark Box
Elevated Plus Maze
Accelerating Rotarod
Cued and Contextual Fear Conditioning
Morris Water Maze
Open Field
total activity, discrete movements, distance traveled rearing
Light Dark Box
• Conflict between
novelty exploration
and avoidance of light
open spaces
• Strain variations
• Responsive to
anxiolytics
Elevated Plus Maze
•Exploration vs. aversion to
open space - prefers dark
enclosed arms
•Score entries into arms over 5
min trial
•Anxiolytics increase entries into
open arms - widely used in drug
discovery
PICTURE CREDITS: EUROPEAN MOLECULAR BIOLOGY LABORATORY
Accelerating Rotarod
Cued and Contextual
Conditioning
•Learning and memory test
•Pair footshock with
auditory cue – see freezing
behavior
•Day 2 return to same
chamber - Freezing is a
measure of contextual fear
•Move to different chamber,
test freezing (control for
context)
•Give auditory cue in
different chamber –
freezing is measure of cued
fear
Morris Water Maze
Opaque water bath
Containing hidden
platform
Visual cues
Computer image
Probe Trial
Acknowledgements
•
•
•
•
Jacqueline Crawley
Christian Newcomer
Michela Gallagher
Nadine Forbes
Questions?