Biology Reference
In-Depth Information
Table 1
Scoring table for the assessment of cold allodynia
Response
Score
No response
0
Startle response lasting less than 1 s
1
Clear withdrawal lasting between 1 and 5 s
2
Clear withdrawal lasting between 5 and 30 s (with or without licking)
3
Clear withdrawal lasting over 30 s (with or without licking and repeated shaking)
4
3. When a response is observed in the form of an acute with-
drawal upon stimulation at any point during stimulation, this
is noted and the paw will no further be stimulated with the
same filament or with a filament of a higher force.
4. When no response is observed, continue with the next filament.
Continue increasing the filament and repeat until the animal
responds. This response is noted.
Repeat the entire testing sequence to obtain results in duplex.
1. Spray 20
l of acetone in one fluent application on the plantar
surface by using the 1 ml syringe with bent needle.
2. Observe the response of the animal and score according to the
scoring table (Table
1
).
3. After 2 min rest, repeat the sequence to obtain results in
duplex.
μ
3.3 Assessment
of Pain: Cold Allodynia
3.4 Treatment
with ARA290
1. Make stock solution of ARA290 of 1 mg/ml in PBS and store
at 4°C.
2. Administer 30
μ
g/kg ARA290 or vehicle (PBS) in a total vol-
ume of 200
l intra peritoneally (i.p.) with a 1 ml syringe
mounted with a 25 G needle (see Note 4).
μ
Sixteen animals were given the spared nerve injury as previously
described and were randomly allocated to a treatment group. Eight
animals received a sham operation. In short, animals were anesthe-
tized with sevoflurane (6% induction, 3% maintenance) and the
trifurcation of the nerve was exposed. No ligation and transection
was performed and the wound was closed in two layers. Twenty-
four hours post injury animals received treatment with ARA290 or
vehicle 5 times at 2 day intervals followed by once a week maintenance
therapy. Within the first 2 weeks following nerve injury, vehicle-
treated animals showed rapid development of tactile allodynia to
the lowest applicable force of 0.004 g. In contrast, i.p. injections
with ARA290 produced long-term relief of tactile allodynia lasting
3.5
Typical Results
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