Project
protocol
-
Contents
Workflow
and
sampling
Step |
Procedure |
Equipment |
Data
collected |
1 |
Elevated
maze
test
(5
min) |
Elevated
maze |
Time
spent
in
open
arms,
enclosed
arms,
middle
platform,
and
total
number
of
entries
into
the
arms |
2 |
Open
field
test
under
dim
lighting
conditions
(5
min) |
Opto-Varimex |
Distance
traveled,
ambulatory
time
in
the
horizontal
plane,
and
number
of
vertical
movements
(rearings) |
3 |
Open
field
test
under
bright
lighting
conditions
(5
min) |
Opto-Varimex |
Distance
traveled,
ambulatory
time
in
the
horizontal
plane,
and
number
of
vertical
movements
(rearings) |
4 |
Novelty-induced
fear
test
(10
min) |
Opto-Varimex |
Number
of
approaches
to
food,
latency
to
start
eating,
and
time
eating |
5 |
Plasma
corticosterone
measured |
Restraining
cages |
Plasma
corticosterone
levels |
Equipment
and
supplies
- Elevated
maze:
constructed
entirely
of
black
Plexiglas,
consisting
of
two
open
arms
(10
x
5
cm)
and
two
enclosed
arms
(30
x
5
x
15
cm)
extending
from
a
center
platform
(5
x
5
cm);
a
0.5
cm
high
Plexiglas
strip
is
attached
to
the
open
arms
of
the
maze
to
prevent
animals
from
falling;
maze
is
elevated
31
cm
off
the
floor;
testing
performed
in
a
windowless
room
under
dim
light
(two
fluorescent
15
W
bulbs
directly
facing
the
wall
on
a
shelf
4
m
away
and
1
m
above
the
center
of
the
maze)
- Opto-Varimex
computerized
activity
monitor
(Columbus
Instruments,
Columbus
OH)
under
dim
lighting
conditions
(described
above)
or
bright
lighting
conditions
(two
fluorescent
15
W
bulbs
located
70
cm
over
the
center
of
the
apparatus)
- Lucite
restraining
cages
(Harvard
Instruments)
Reagents
and
solutions
- Ethanol/water
(50%
v/v)
for
wiping
down
each
apparatus
between
tests
for
each
mouse
Acclimation
period
One
week
after
arrival
at
the
test
facility,
mice
were
moved
from
the
housing
colony
room
to
the
testing
area
in
their
home
cages
and
allowed
to
adapt
to
the
new
environment
and
light
level
for
1
h
before
testing.
Procedure:
Elevated
maze
test
- Group
1:
naive
mice
- Scoring
is
for
5
min
under
dim
light.
- Each
mouse
is
placed
at
the
intersection
of
the
four
arms
so
that
its
head
is
in
the
center
of
the
platform.
- Mice
are
considered
to
be
in
open
or
closed
arms
only
when
they
have
all
four
legs
within;
otherwise
they
are
scored
as
being
on
the
middle
platform.
- Parameters
are
recorded
using
an
Apple
IIe
computer.
- Percentage
of
time
spent
in
the
open
arms
is
calculated
over
the
sum
of
time
spent
in
the
open
and
closed
arms;
the
percentage
of
entries
into
the
open
arms
is
calculated
based
on
the
total
arm
entries.
Procedure:
Open
field
test
- Group
1:
same
animals
tested
in
Elevated
Maze
one
day
prior;
under
dim
lighting
conditions
- Group
2:
naive
mice;
under
bright
lighting
conditions
- Mice
are
placed
in
the
center
of
the
Opto-Varimex
open
field
arena
and
spontaneous
activity
scored
for
5
min
with
a
computerized
activity
monitor.
Procedure:
Novelty-induced
fear
test
- Group
3:
naive
mice;
under
bright
lighting
conditions.
- Mice
are
food-deprived
for
24
h.
- Mice
are
placed
in
the
corner
of
the
Opto-Varimex
open
field
that
contains
a
pellet
of
rat
chow
in
the
center.
- Scoring
is
for
10
min.
Procedure:
Plasma
corticosterone
levels
- Group
4:
naive
mice;
no
stress
- Group
5:
naive
mice;
30
min
of
restraint
stress
- For
determination
of
basal
corticosterone
values,
mice
are
removed
from
their
home
cage,
and
rapidly
transferred
to
a
different
room
where
trunk
blood
is
collected
(<
1
min).
- For
determination
of
stress-induced
corticosterone
values,
mice
are
placed
in
individual
lucite
restraining
cages
for
30
min;
after
stress
period,
mice
are
rapidly
transferred
to
a
different
room
and
trunk
blood
collected.
- Basal
and
stress
groups
are
performed
concurrently.
- Mice
are
euthanized
in
a
predetermined
order,
randomized
for
both
strain
and
treatment.
- Plasma
corticosterone
levels
are
determined
by
radioimmunoassay.
Data
collected
by
investigator
- Elevated
Maze
- time
spent
in
open
arms
- time
spent
in
closed
arms
- time
spent
in
middle
platform
- total
number
of
entries
into
the
arms
- Open
Field
Test
- distance
traveled
- ambulatory
time
- vertical
movements
- Novelty-induced
Fear
Test
- number
of
approaches
to
food
- latency
to
start
eating
- time
eating
- Plasma
Corticosterone
Levels
- basal
level
- stress-induced
level
|