Arginine vasotocin promotes urea permeability through urea transporter expressed in the toad urinary bladder cells

We previously isolated a cDNA of a urea transporter (Bufo UT) from the kidney of the marine toad, Bufo marinus, and demonstrated that the Bufo UT was specifically localized on the epithelial membrane of the early distal tubules in the kidney and urinary bladder. In the present study, the function of Bufo UT was investigated using a Xenopus oocytes expression system. Further, we examined the effects of arginine vasotocin (AVT) on urea transport in isolated cells from the toad urinary bladder. When expressed in Xenopus oocytes Bufo UT induced more than a 10-fold increase in [(14)C]urea uptake compared with water-injected control oocytes. Phloretin, a urea transport inhibitor, fully blocked the increase of urea uptake. In epithelial cells isolated from the toad urinary bladder, addition of AVT to the medium increased the urea uptake in a concentration-dependent manner (10(-12)-10(-8)M). To examine the relationship between the Bufo UT protein expression and an increase of urea transportability, we analyzed the time course of the Bufo UT expression levels and urea uptake in the cells treated with 10(-8)M AVT. Treatment of 10(-8)M AVT increased the urea uptake in the cells after 24 and 48h incubation, but not after 12h. According to the immunoblot analysis, UT protein expression was coincident with the results of urea uptake in the AVT-treated cells. These results suggest that Bufo UT isolated from the kidney, functions as an AVT-mediated urea transporter in the urinary bladder of the toad.