Enhanced Cholesterol Diet effects Epithelial Na+ Channel levels

The epithelial Na+ channel (ENaC) is a trimeric transmembrane channel expressed in multiple secretory organs.  ENaC is responsible for regulation of Na+ transport and balance in the kidney. The cellular effects of regulating membrane cholesterol and fluidity of ENaC activity are known, however little has been sufficiently described concerning its regulation in vivo. Using a rat model of metabolic syndrome, we use an Otsuka Long Evans Tokushima Fatty (OLETF) rat as a suitable animal model of type II diabetes with obesity. Also, we use lean control rats (LETO) that are supplemented with a high cholesterol diet (2%) (HCD). Furthermore, we treat these certain samples for 16 weeks with Angiotensin II receptor blocker (ARB), which is medication that blocks the action of angiotensin II by preventing angiotensin II from binding to angiotensin II receptors on blood vessels. In addition, we use a group of OLETFs in which ARB was removed 5 weeks before the end of the study, to analyze the long-term effects of ARB. The methodology we use consists of protein assay, western blotting, and desitometry. Protein assay will help us accurately identify the exact amount of protein within each kidney sample. Western Blotting is a technique best used for determining the molecular weight of proteins; followed by desitometry, which is used to record the correct weight for each band that appears on the membrane of the western blot. The data should suggest a cellular effect of modifying membrane cholesterol and fluidity on the ENaC activity in the kidney.