Animal Behavior Analysis Solutions

Special
Behavior

Paw Licking in rats

The LABORAS system

∇ Metris has released a new module to recognise paw licking

in rats. This behaviour is characterised by licking of the hind
limb and is an important parameter in nociceptive tests.

∇ Engineers at Metris developed an algorithm that assesses

paw licking from the LABORAS sensor signals.

∇ Paw licking is an important behaviour in the research for

analgesic treatments.

∇ LABORAS paw licking provides a fast and reliable

assessment of analgesic effects of compounds.

For commercial information please contact sales@metris.nl.

Metris BV
Kruisweg 801a • 2132 NG Hoofddorp • The Netherlands • Tel. +31 (0)23 562 3400 • www.metris.nl • info@metris.nl
 Animal Behavior Analysis Solutions

Paw Licking in rats

Metris b.v. has released a new
LABORAS module to automatically
record and analyse the paw licking
response after local injection with
formalin into one of the hind limbs. This
module replaces and standardizes the
traditional time-consuming human
observations. Researchers conclude
that this new paw licking algorithm
provides a fast and reliable
measurement for the assessment of
analgesic effects of compounds.
doses of four reference analgesic compounds
(morphine, MK-801, lacosamide, pregabalin).
RESULTS
Functional validation
No significant difference could be detected
TM
between the LABORAS paw licking module
and the mean of the three human observers
(Figures 1, 2, 3 4 and 5).
Pharmacological validation
TM
The LABORAS paw licking module revealed
ED50 values that were in accordance with
previous findings.

INTRODUCTION

Metris b.v. in cooperation with Gedeon Richter 140

Ltd. Neuropharmacology Department 120

developed new software to automatically 100
obs1
obs2
measure the pain-related behavior of rats in
licking time (s)

LABORAS
80
the so-called formalin test. LABORAS™ is able obs3

to automatically analyze several different 60 mean of obs

behaviors of rats and mice (locomotion, 40

immobility, rearing, grooming and additionally 20

eating and drinking). The formalin add-on *
0
software module (algorithm) was specially 1 2 3 4 5 6 7 8 9 10 11 12
designed to automatically detect the paw 5 min periods

licking response of rats after formalin injection

Fig. 1 Comparison between LABORAS™
in the right hind limb. The goal of this special
formalin test and observers’ scores after
software is to replace the traditional and time
pretreatment with saline (n=3-3).
consuming observational work by humans.

To verify the performance of the formalin

response detection algorithm a special 160
experiment was prepared. During the 140
acquisition with the LABORAS™ system a 120
video camera recorded the rat at the side of obs1
licking time (s)

100 obs2
platform. After acquisition, LABORAS™ LABORAS
80
automatically analyzed the behavior of the obs3
60
animals and classified paw licking with the new mean of obs

formalin algorithm. The video recording of the 40

measurement was independently scored by 20

three observers. The results of the 0

1 2 3 4 5 6 7 8 9 10 11 12
LABORAS™ analysis and the observers’ 5 min periods
scores were compared.
Fig. 2 Comparison between LABORAS™
The LABORAS™ paw licking module was formalin test and observers’ scores after
pharmacologically validated by using different pretreatment with vehicle (n=3-3)

Metris BV
Kruisweg 801a • 2132 NG Hoofddorp • The Netherlands • Tel. +31 (0)23 562 3400 • www.metris.nl • info@metris.nl
Metris
 Animal Behavior Analysis Solutions

DISCUSSION AND CONCLUSIONS

120

100
It is well established in the literature that the
obs1
obs2
formalin paw licking response curve is
80
biphasic. After formalin injection an increased
licking time (s)

LABORAS

60
obs3
time of licking can be measured until 5 or 10
mean of obs
minutes which is called phase 1. Between 10
40
and 15 min licking behavior almost completely
20 disappears, and after 15 min following formalin
injection phase 2 can be observed wherein
0
1 2 3 4 5 6 7 8 9 10 11 12
paw licking is increased again up to 30-35 min.
5 min periods Our results are in correspondence with these
previous findings. Either LABORAS formalin
Fig. 3 Comparison between LABORAS™ measurement or observers’ scores resulted in
formalin test and observers’ scores after the biphasic formalin response curve
pretreatment with treatment A (n=3-3) mentioned above.

When evaluating the LABORAS™ formalin

result and observers’ scored data we can find
100
that the difference between observers and
90
80
obs1 LABORAS is not higher than can be measured
obs2
70
between observers. The highest difference can
LABORAS
licking time (s)

60 obs3 be found at the descending part of 2nd phase.

50 mean of obs This is the part of pain related licking behavior
40 where animals are licking their paw frequently
30
* for a short time. There is a possibility of a small
20 systematic error made by LABORAS™ or by
10
observers, as well. Probably, the longer
0
1 2 3 4 5 6 7 8 9 10 11 12
reaction time of humans can make such a
5 min periods difference. The difference is small and does
not disturb reliable assessment of inhibition by
Fig. 4 Comparison between LABORAS™ a drug which was the main goal of the test.
formalin test and observers’ scores after
pretreatment with treatment B (n=3-3) Dr. Kristina Kostás et al.1, Gedeon
Richter Ltd., Dept. Neuropharmacology,
Budapest, Hungary:
120

100 Laboras Our conclusion is that the LABORAS™

obs1 paw licking software provides a fast
licking time (sec)

80
obs2
and reliable measurement to assess the
60
obs3
effects of analgesic compounds. The
mean of obs
40
ED50 values measured with the
20 LABORASTM paw licking module are in
0
agreement with previous findings.
0 5 10 15 20 25 30
5 min periods Reference
1. Kordás KS, Galgóczy K, Bulthuis RJA, Schekkerman A,
Fig. 5 Comparison between LABORAS™ Horváth C. New automated measurement of formalin
formalin test and observers’ scores after induced paw licking in rats by LABORASTM system. Poster
pretreatment with treatment C (n=3-3) at International Forum on Pain Medicine, Sofia, Bulgaria,
2005.

Metris BV
Kruisweg 801a • 2132 NG Hoofddorp • The Netherlands • Tel. +31 (0)23 562 3400 • www.metris.nl • info@metris.nl
 Animal Behavior Analysis Solutions

Metris BV
Kruisweg 801a • 2132 NG Hoofddorp • The Netherlands • Tel. +31 (0)23 562 3400 • www.metris.nl • info@metris.nl