Crabbe5
Protocol
Project
protocol
—
Contents
Workflow,
sampling,
and
experimental
treatment
Equipment
Reagents,supplies,
and
solutions
Procedure:
Parallel
rod
floor
test
for
motor
incoordination
Data
References
Strains
were
serially
tested
on
four
floor
types
(see
details
below).
Animals
were
randomized
into
two
groups
per
strain.
Group
1
floor
order:
E,
A,
D,
B
Group
2
floor
order:
A,
E,
D,
B
Each
mouse
was
tested
before
(saline
control)
and
after
EtOH
treatment.
Each
mouse
was
tested
on
four
platforms
of
the
parallel
rod
floor
apparatus
(see
Equipment
below)
on
a
rotating
3-day
injection
schedule.
Mice
were
undisturbed
for
six
days
between
platforms.
Workflow
| Day |
Injection |
Tested |
Data
collected |
1 |
Saline |
Yes |
Yes |
2 |
Saline |
Yes |
Yes |
3 |
EtOH |
Yes |
Yes |
4-9 |
None
* |
No |
No |
*
Recovery
period
before
next
platform
Treatment
| Dose |
2g
EtOH
per
kg
body
weight
[g/kg] |
| Delivery |
intraperitoneal
injection
(i.p.) |
Equipment
•
Parallel
rod
floor
apparatus
is
described
in
detail
in
Crabbe
et
al.,
2003
and
more
information
in
Kamens
et
al.,
2005.
Briefly,
a
clear
acrylic
box
sits
is
placed
on
top
of
a
parallel
rod
floor
which
is
1
cm
above
a
base
plate.
The
apparatus
is
set
inside
an
activity
monitor.
The
activity
monitor
is
housed
in
a
sound-insulated
black
acrylic
chamber
equipped
with
fluorescent
lighting
and
a
ventilating
fan.
Errors
are
recorded
by
an
Apple
IIE
computer
when
a
paw
slips
through
the
rods
and
completes
an
electrical
circuit
with
the
base
plate.
The
activity
monitor
records
horizontal
distance
traveled
(detected
by
photocell
beam
breaks).
Parallel
rod
floor
configurations
| Floor
*
(apparatus) |
Rod
diameter
(mm) |
Inter-rod
distance
(mm) |
A |
1.6 |
4 |
B |
1.6 |
6 |
D |
3.2 |
6 |
E |
4.8 |
6 |
*
A,
B,
D,
E
are
described
in
Kamens
et
al.,
2005.
Configuration
C
was
not
used
for
testing.
•
AccuScan
Activity
Monitor,
Columbus
OH,
USA
Reagents,
supplies,
solutions
- Test
agent:
ethanol
(EtOH),
Pharmoco
Inc,
Brookfield
CT,
USA;
- EtOH
solution
prepared
at
20%
v/v
in
saline
- Saline
(physiological
saline):
0.9%,
Baxter
Healthcare
Corp,
Deerfield
IL,
USA
- Isopropyl
alcohol:
10%
for
wipe
downs
between
testsAcclimation
to
test
conditions
Acclimation
Mice
are
acclimated
to
room
environment
45-60
min
before
testing.
Procedure:
Parallel
rod
floor
test
for
motor
incoordination
a.
Randomly
selected
mice
are
i.p.
injected
with
saline
or
EtOH.
b.
Each
mouse
is
immediately
positioned
in
the
center
of
the
parallel
rod
floor
apparatus
(see
Equipment
above
for
details),
and
testing
begins.
c.
Mice
are
monitored
and
data
collected
over
15
min
of
testing.
d.
The
apparatus
is
cleared
and
wiped
with
10%
isopropyl
alcohol
after
each
test.
Data
collected
by
investigator
Distance
traveled
and
number
of
paw
slips
(errors)
Definitions
&
formulas
Motor
incoordination
(err)
=
(#errors
/
distance
traveled)
x
100
|
|
Test age:
7-14wks
