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Am J Physiol Renal Physiol 283: F1337-F1350, 2002. First published July 24, 2002; doi:10.1152/ajprenal.00165.2002
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Vol. 283, Issue 6, F1337-F1350, December 2002

Electrolyte and fluid secretion by cultured human inner medullary collecting duct cells

Darren P. Wallace1,2,3, Marcy Christensen1, Gail Reif1, Franck Belibi1, Brantley Thrasher4, Duke Herrell4, and Jared J. Grantham1,2,3

1 Kidney Institute and Departments of 2 Biochemistry and Molecular Biology, 3 Medicine, and 4 Urology, University of Kansas Medical Center, Kansas City, Kansas 66160

Inner medullary collecting ducts (IMCD) are the final nephron segments through which urine flows. To investigate epithelial ion transport in human IMCD, we established primary cell cultures from initial (hIMCDi) and terminal (hIMCDt) inner medullary regions of human kidneys. AVP, PGE2, and forskolin increased cAMP in both hIMCDi and hIMCDt cells. The effects of AVP and PGE2 were greatest in hIMCDi; however, forskolin increased cAMP to the same extent in hIMCDi and hIMCDt. Basal short-circuit current (ISC) of hIMCDi monolayers was 1.4 ± 0.5 µA/cm2 and was inhibited by benzamil, a Na+ channel blocker. 8-Bromo-cAMP, AVP, PGE2, and forskolin increased ISC; the current was reduced by blocking PKA, apical Cl- channels, basolateral NKCC1 (a Na+-K+-2Cl- cotransporter), and basolateral Cl-/HCO<UP><SUB>3</SUB><SUP>−</SUP></UP> exchangers. In fluid transport studies, hIMCDi monolayers absorbed fluid in the basal state and forskolin reversed net fluid transport to secretion. In hIMCDt monolayers, basal current was not different from zero and cAMP had no effect on ISC. We conclude that AVP and PGE2 stimulate cAMP-dependent Cl- secretion by hIMCDi cells, but not hIMCDt cells, in vitro. We suggest that salt secretion at specialized sites along human collecting ducts may be important in the formation of the final urine.

kidney; chloride transport; salt secretion


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