Objective Quantitation of Focal Sweating Areas Using a Mouse Sweat-assay Model

Abstract

In vivo sweat quantitation assays are required for the development of drugs for the management of focal hyperhidrosis before clinical trials; however, in vivo assays, particularly mouse models, are rare. Even in sweat assays using mice, sweating is quantitated by manually counting the number of sweating spots, which can contribute to various errors owing to arbitrary judgment. In this study, we developed a mouse sweat-assay model and a method for quantitating the amount of sweating to remove possible errors. The use of the iodine-starch test in the castor oil-covered hind footpad skin of anesthetized mice resulted in the sweating area being stained blue-black. After the anesthesia and treatment with drugs (pilocarpine, glycopyrrolate, botulinum neurotoxin, myricetin, and myricetin-loaded lipid nanoparticles), the remaining area of the footpad skin was eliminated from the acquired footpad images using ImageJ. Blue pixels extracted from the footpad image are automatically adjusted using the Phansalkar method, where the percentage of the blue area was determined based on the whole hind footpad skin area, finally indicating the percentage of the sweating area. Using this mouse model and analysis for sweat assays, a clear difference between the control group and antiperspirant-administered group was observed with respect to the sweating area % with no error. In conclusion, this assay can be used as a preclinical tool to screen potential antiperspirant drugs. Graphic abstract: Overview of the mouse-model sweat assay and objective quantitation of the focal sweating area.

Keywords: Mouse hind footpad perspiration; Mouse model; Non-arbitrarily judged quantitation; Sweat assay; Sweat quantitation.