Na-K-Cl cotransport in apical membrane of rabbit renal papillary surface epithelium

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Abstract
The renal papillary surface epithelium is exposed to pelvic urine on its apical surface and to inner medullary interstitium on its basolateral surface. To investigate transport in this epithelium, we dissected it free from the renal papilla of rabbits and mounted it in a chamber that allowed both sides to be bathed independently. Cell volume was measured at 25 degrees C utilizing computerized quantitative microscopy. Addition of ouabain (10(-4) M) to the basolateral solution induced a 20% volume increase. This volume increase was completely inhibited by the removal of apical bath NaCl, Na+, K+, or Cl- but not by the removal of urea. Bumetanide, down to 10(-9) M in the apical bath, completely inhibited the ouabain-induced swelling. Changes in apical bath osmolality, resulting from addition or removal of NaCl, caused cell volume changes that were greater than could be accounted for by osmotic water flow alone. This hyperresponse was blocked by bumetanide and was stimulated by vasopressin (10(-8) M). These observations are consistent with the presence of Na-K-ATPase in the basolateral membrane and a bumetanide-sensitive, vasopressin-responsive Na-K-Cl co-transporter in the apical membrane.