Use of proteases for the evaluation of the different adiponectin isoforms in the dog

Abstract

Adiponectin (ADP) is an adipokine secreted by adipose tissue with anti-inflammatory, antiatherogenic, and antidiabetic properties. In human serum, it is presented as three different forms: low molecular weight (LMW), medium molecular weight (MMW), and high molecular weight (HMW). High molecular weight isomer is the most active form of ADP and is more closely related with obesity-induced insulin resistance and metabolic syndrome than total ADP. Selective protease treatment can be used in humans to isolate the different ADP isoforms but this has not been applied in any veterinary species. Therefore, the objective of this study was to evaluate if the selective protease digestion is able to differentiate serum ADP isomers in dog samples, and if these isomers could change in obese dogs after a weight loss program. A Western blotting analysis confirmed that digestion with protease K showed only the HMW forms of ADP, whereas the use of protease A showed the HMW and MMW forms. This specific protease digestion was applied to serum obtained from 14 obese beagle dogs before and after a weight loss program and total ADP, HMW, and LMW forms increased significantly after the weight reduction. In conclusion, the use of selective protease digestion can be applied in canine serum as a procedure for detecting the different ADP isomers. In addition, by this procedure, it was showed that the HMW and LMW forms were increased after a weight loss program in our experimental conditions.

Keywords: Dogs; HWM; Proteases; Serum; adiponectin.