Uroplakins do not restrict CO 2 transport through urothelium

Abstract

Lipid bilayers and biological membranes are freely permeable to CO 2, and yet partial CO 2 pressure in the urine is 3-4-fold higher than in blood. We hypothesized that the responsible permeability barrier to CO 2 resides in the umbrella cell apical membrane of the bladder with its dense array of uroplakin complexes. We found that disrupting the uroplakin layer of the urothelium resulted in water and urea permeabilities (P) that were 7- to 8-fold higher than in wild type mice with intact urothelium. However, these interventions had no impact on bladder P CO2 (∼1.6 × 10 -4 cm/s). To test whether the observed permeability barrier to CO 2 was due to an unstirred layer effect or due to kinetics of CO 2 hydration, we first measured the carbonic anhydrase (CA) activity of the bladder epithelium. Finding none, we reduced the experimental system to an epithelial monolayer, Madin-Darby canine kidney cells. With CA present inside and outside the cells, we showed that P CO2 was unstirred layer limited (∼7 × 10 -3 cm/s). However, in the total absence of CA activity P CO2decreased 14-fold (∼ 5.1 × 10 -4 cm/s), indicating that now CO 2 transport is limited by the kinetics of CO 2 hydration. Expression of aquaporin-1 did not alter P CO2 (and thus the limiting transport step), which confirmed the conclusion that in the urinary bladder, low P CO2 is due to the lack of CA. The observed dependence of P CO2 on CA activity suggests that the tightness of biological membranes to CO 2 may uniquely be regulated via CA expression. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.