Nociceptive Withdrawal Responses 1

The Effect of Tail Position on the Nociceptive Withdrawal Response of Rats

Sarah Donaire, Rebecca Mullins, and Jenny Yang
Massanutten Regional Governors School
December 20, 2013














Nociceptive Withdrawal Responses 2

In past years, many tests have been conducted on various organisms to understand
reflexes and movement (Camhi, 1980; Cleland and Bauer, 2002; Kim, et al., 2006) . As
technology improved and became more advanced, scientists began to uncover connections
between the reflexes they were researching and the nervous system. Further discoveries have
contributed to advances in societys understanding of reflexes and the way that animals, and
even humans, transmit signals and respond to stimuli (Camhi 1980). Our experiment involves
nociceptive withdrawal testing on rats and is different, because the rat is placed with its tail in
seven different locations around its body. These tests show not only how heat stimulus affects
withdrawal responses, but also how difference in initial tail position can influence certain
reflexes.
A little background information is needed for a reader to understand our
research. Reflexes are automatic, involuntary responses to stimuli. The stimulation of a muscle
or nerve can produce many different reflex responses, depending on the location and original
position of the stimulated nerve (Cleland and Bauer, 2002). This is to help the animal get away
from predators or any other types of danger it may face in its natural habitat (Camhi 1980).
The involuntary escape response in cockroaches, from an air stimulus, has been studied
(Camhi 1980). Scientist examined how cockroaches would respond when stimulated with a puff
of air, and how the direction of the stimulus related to the direction of the escape response. It
was observed that the puff of air causes cockroaches to begin to run, but that response is absent if
the two posterior sensory appendages (known as cerci) are removed (Camhi 1980). The
underside of the cerci contain around two hundred tiny hairs, which move when air is blown and
therefore cause the resulting reflex of the cockroach. Scientists covered these hairs with wax,
and then puffed air onto the cockroach once again. Without the movement of the hairs on the
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cerci, the response was absent. Scientists then used toads to further test this (Camhi 1980). They
put a cockroach and a toad in a small container to simulate what it would be like in nature. Fifty
five percent of the control cockroaches that had nothing done to their cerci escaped the toad,
forty seven percent of the cockroaches that had just their abdomen covered with wax escaped
and only eight percent of the insects that had their cerci covered got away (Camhi, 1980). This
showed that these insects need the cerci to be able to sense movement of air and effectively
escape predators. In conclusion, for the majority of the cockroaches that had nothing done to
their cerci, they responded with a movement away from the stimulus whether it be a live toad or
a puff of air (Camhi 1980).
In mammals, the escape response is replaced with a nociceptive withdrawal
response. The nociceptive withdrawal response is the bodys response to stimulation that could
possibly be damaging. The area stimulated withdraws from from potentially dangerous stimuli
(Bear, et al., 2007).
Years ago, an extensive study on the nociceptive withdrawal reflexes of cats and dogs
took place (1910). Scientists wanted to determine how the receptive field related to the
underlying muscle tissue of limbs. The term receptive field is applied to a series of receptive
points, that when stimulated together can produce a specific reflex movement, such as a
dog scratching his ear with his hind limb (Sherrington, 1910). The animals underwent either a
decerebrate (removal of the cerebrum) or decapitate (removal of everything above the brainstem)
procedure, then nerves on the hind legs were tied and stimulated mechanically, and the resulting
movement was recorded. Flexion and extension of limbs was easily observed in both
decerebrate and decapitate preparations (Sherrington, 1910). A joint and the adjacent muscles
bending, like a pair of scissors snapping shut, is known as flexion. When the opposite is
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happening, and a muscle is being opened or stretched, it is called extension (Bear, et al., 2007).
However, the relaxation of limbs could not be observed unless the limb was rigid. To produce
this background of rigidity on a decapitate preparation, the opposite limb was stimulated,
producing an extension in the leg being tested. The decerebrate preparation produced a natural
rigidity of limbs in which relaxation of muscles could easily be seen, which made it much more
convenient and effective than the decapitate preparation (Bear, et al., 2007). It was observed that
the receptive field does not only consist of an area on the surface of the skin, but also includes
and takes into account the thickness of the limb and the muscles and veins inside. Therefore,
when large nerve trunks of limbs were stimulated, the results from each of them were flexion,
and practically the same (Sherrington, 1910).
In the late 1990s, a group of scientists found evidence that contradicted Sherringtons
conclusions. The scientists used rats to determine whether a separate reflex field exists for each
muscle, or if the fields are more interconnected (Schouenborg, 1990). Some rats were
spinalized, (the spinal cord was detached between the front and hind limbs) while others spinal
cords were kept intact. A CO2 laser was triggered at different areas of the rats hind limbs, and
resulting movements were recorded. It was observed that a few hours after spinalization
occurred, the receptive field directly corresponded to the area of muscle stimulated. The skin
area most effectively withdrawn corresponded to the most sensitive area of the receptive field. It
was concluded that there is a separate reflex path to each muscle, each of which have a well
organized receptive field. These findings contradict Sherringtons conclusion, which shows that
more research is needed to gain a correct understanding of receptive fields (Schouenborg, 1990).
In a study by Dr. Corey L. Cleland and Dr. Ross E. Bauer, the experimental conclusions
of Sherrington and Schouenborg were hybridized. Rats tails were stimulated and the trajectory
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of the spatial response movement was observed. Like in Schouenborgs experiment, both
spinalized and intact rats were used. The rats were placed in a tube and held upright, while the
tail was stimulated with a small pinpoint of heat. The direction and strength of the tail
movement observed and analyzed. The experiment showed that a spinalized and intact rats
showed little difference, except when the tail was stimulated dorsally and ventrally (Cleland and
Bauer, 2002). In spinalized rats, when heat was applied to the ventral area, instead of showing
movement away from the heat source the tail moved toward the stimulus. In intact rats, the
opposite was true; all stimulation points caused the tail to move away except for when the dorsal
area of the tail was tested (Cleland and Bauer, 2002).
Experiments have been conducted on animals that concluded that nociceptive reflexes are
influenced by initial position of the body part being tested on. In 2006, an experiment conducted
at the Rehabilitation Institute of Chicago furthered the research of the effect initial position of a
body part has on its reflex. Specifically, the purpose of the research was to observe the influence
of hip proprioceptors by nociceptive electrical stimulation of the foot in subjects with chronic
spinal cord injury (Kim, et. al. 2006). It was hypothesized that the change of the position of hip
would influence excitability of the spinal flexor reflexes. Ten subjects with histories of spasms
were chosen. A patients foot was secured and stimulated electrically. Hip torque, position, and
velocity were measured. This was done with the hip positioned three different ways, and also
with the hip at midposition during hip flexion and extension movement. It was concluded that
ankle and hip torques were significantly affected by hip position, with the strongest response
following the hip extended position, rather than the flexed position (Kim, et. al. 2006).
Movement of the hip produced a larger reflex response to electrical stimulation than isometric
conditions. This difference in movement conditions was statistically significant. These results
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were generally constant. These results suggest that both static and dynamic hip proprioceptor
components affect withdrawal reflexes. It was also concluded that static components from hip
proprioceptors enhanced responses to electric stimuli, and that imposed movement of the hip
promotes responses regardless of movement direction (Kim, et. al. 2006).
In 2013, the Rehabilitation Institute of Chicago conducted another experiment on the
effect initial position of a body part has on its reflex. The researchers focused on the withdrawal
reflexes in the upper limb since they have not been examined to the same extent as the lower
limbs. It was still unclear if the withdrawal reflex in the lower limb was also present in the upper
limb. The purpose of this research was to determine if the withdrawal reflex adapts to different
stimulus locations and postures in the upper limb (Peterson, et. al. 2013). It was hypothesized
that in all conditions the withdrawal reflex would be to move the limb away from the stimulus.
Twenty-two healthy adults with no history of upper limb impairment were chosen for this
research. The subjects were seated in a chair with their right forearm secured in a plastic cast. At
different points in the experiment their right thumb, index finger, and pinky finger were
stimulated electrically. Each of these fingers was stimulated with the arm in three different
postures: flexed, mid-range, and extended. Direction, magnitude of the reflex force, and the
reflex latencies were measured from all three postures and digits. The direction of the endpoint
force elicited by the withdrawal reflex varied with changes in the arm posture (Peterson, et. al.
2013). The researchers found that the responses were significantly stronger in the flexed posture
than the middle or extended posture. It was also found that the magnitude of the force response
from the withdrawal reflex was greatest with stimulation of digit V (the pinky finger) compared
to the other two digits. Both of these result were generally constant throughout their research.
These findings supported their hypothesis that the withdrawal reflex in the upper limb was to
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move away from stimulation. As for the direction of the withdrawal response, it was found to
depend on the posture of the arm (Peterson, et. al. 2013).
A 2013 studys results contrast with the two experiments above. The research tested the
adaptability of the human nociceptive withdrawal reflex; that is, the the mechanism to withdraw
a limb from a potentially dangerous situation. It was unknown whether this reflex in the human
upper limb is adaptable based on position, or if it could simply be modified by short-term
practicing of an opposing motor action (Eckert and Riley, 2013). Ten adults participated in this
study; their arms were placed in five static positions and stimulated with an electric shock, and
the nociceptive withdrawal responses were examined. The adults were also subjected to
conditioning tasks, where they were asked to extend their arm against an electrical
stimulation. Then the trials were repeated, and the withdrawal responses were recorded. It was
observed that different arm positions were independent, and therefore any arm position could be
chosen at random in comparison studies (Eckert and Riley, 2013). It was concluded that
changing the elbow and shoulder position results in a nociceptive withdrawal response of the
same receptive field, and therefore the joint position does not affect withdrawal reflex. This
conclusion counters the results of the two previously explained studies, as their findings saw that
the nociceptive withdrawal reflex was in fact dependent on position. The conditioning task was
found to cause no change in latency between stimuli given before and after conditioning. These
findings caused scientists to conclude that the nociceptive withdrawal response is resistance to
change, specifically short-term motor practices (Eckert and Riley, 2013).
Our experiment is focused on observing changes in withdrawal responses, if they exist,
when the tail is initially located in seven different positions. The rat is weighed and then the tail
is measured to ensure that the tracking bands are made in the correct locations. After the tail is
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marked the rat is placed into a cylindrical dark tube. The order of tail angles are randomly
selected. The stimulation point on the tail is stimulated fourteen times during an experiment with
at least a four minute interval between each stimulation. An advanced camera is used to track the
points on the tail and follow the rat tails withdrawal path.
Although studies of rat tail reflexes have already taken place (Cleland and Bauer, 2002),
in all these tests, the rat had been contained, forcing it into an unnatural position. Because the rat
was uncomfortable in the upright tube setup, using those results to describe the stimulation
responses for a rat in its natural habitat would be incorrect or misleading. A comfortable rat
would have all four feet and its tail flat on the ground. The tail is flexible and can be in different
placements around the body with the rat in a natural position. The reflex movement of the tail
has never been tested before with the animal in this position. Our research group is expanding on
Dr. Clelands research. The aim of our research is to stimulate a specific point on rats tails,
from either side, with heat from a laser. This point is stimulated while the tail of the rat is in
seven different positions, then examine and analyze the trajectory of the response reflex using
advanced cameras. The results of our experiment can then be compared to the results of Dr.
Cleland and Dr. Bauers experiment, to see how positioning of the animal affects the movement
of rat tails when stimulated.










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Eckert, Nathan and Zachary A Riley. (2013). Adaptability of the Nociceptive Withdrawal
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Kim, Yongchul, et. al. (2006). Modulation of flexor reflexes by static and dynamic hip
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