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  • Development and validation of an automated system for detection and assessment of scratching in the rodent.

Development and validation of an automated system for detection and assessment of scratching in the rodent.

Journal of neuroscience methods (2012-09-14)
Marc Marino, Polly Huang, Shelle Malkmus, Erin Robertshaw, Elaine A Mac, Yuri Shatterman, Tony L Yaksh
ABSTRACT

Pruritus, the sensation of itch, which evokes reflex scratching behavior, has a diverse etiology. Because of its clinical significance, mechanisms of pruriception are an important topic. In the present work we describe and validate a paw motion detector (PMD) system. The system employs a small removable metal band placed on one hind paw that provides a signal indicative of paw movement through perturbation of an electromagnetic (EM) field. C57Bl/6 mice were fitted with a unilateral hind paw band and adapted to testing cylinders equipped with EM signal emission and detection. The following observations were made: (1) in mice, unilateral SQ injection of 48/80 into the dorsolateral aspect of the neck evoked periodic high frequency bursts of scratching at the injected site with the ipsilateral (banded) but not the contralateral (not banded) hind paw. (2) Cross correlation between PMD and human observer counts after SQ 48/80 using the specified computational algorithm revealed a highly significant correlation. (3) SQ histamine and 48/80 over a 1hour interval produced dose dependent scratching, which diphenhydramine dose dependently reversed. Chloroquine scratching displayed an inverse u-shaped dose response curve, which was insensitive to diphenhydramine. (4) SQ 48/80 at intervals over 28 days showed no change in the scratching response within the same cohort of mice. (5) Power analysis showed 40% changes in scratching activity could be detected at the p<0.05 level with groups of 4 mice. These observations indicate that the system described can efficiently define the actions and pharmacology of pruritogenic agents.

MATERIALS
Product Number
Brand
Product Description

Supelco
Diphenhydramine hydrochloride solution, 1.0 mg/mL in methanol (as free base), ampule of 1 mL, certified reference material, Cerilliant®
Supelco
Diphenhydramine hydrochloride, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Diphenhydramine hydrochloride, ≥98% (HPLC)
Diphenhydramine hydrochloride, European Pharmacopoeia (EP) Reference Standard