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Am J Physiol Renal Physiol (June 29, 2004). doi:10.1152/ajprenal.00448.2003
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Submitted on December 22, 2003
Accepted on June 22, 2004

Adrenergic regulation of salt and fluid secretion in human medullary collecting duct cells

Darren P. Wallace1*, Gail Reif2, Anne-Marie Hedge2, J. Brantley Thrasher3, and Paul Pietrow3

1 Kidney Institute, University of Kansas Medical Center, Kansas City, Kansas, USA; Department of Internal Medicine, University of Kansas Medical Center, Kansas City, Kansas, USA
2 Kidney Institute, University of Kansas Medical Center, Kansas City, Kansas, USA
3 Department of Urology, University of Kansas Medical Center, Kansas City, Kansas, USA

* To whom correspondence should be addressed. E-mail: dwallace{at}kumc.edu.

Transepithelial salt and fluid secretion mediated by cAMP in initial inner medullary collecting ducts (IMCDi) may be important for making final adjustments to the urine composition. We examined in primary cultures of human IMCDi cells the effects of adrenergic receptor (AR) agonists and antagonists on intracellular cAMP levels, short-circuit current and fluid secretion. Epinephrine (1 µM), norepinephrine (1 µM) and isoproterenol (10 nM) individually increased intracellular cAMP levels 57-, 2-, and 25-fold, respectively, and stimulated short-circuit current (ISC) 3.3-, 2.9-, and 3.4-fold, respectively. {beta}-AR activation increased net fluid secretion by cultured human IMCDi cell monolayers from 0.09 ± 0.04 to 0.26 ± 0.05 µL/h/cm2 and freshly isolated rat IMCDi from 0.02 ± 0.01 to 0.09 ± 0.02 nL/h/mm length. In monolayers, these effects were eliminated by blocking {beta}2-AR, but not {beta}1-AR. Activation of {alpha}2-AR with guanabenz inhibited isoproterenol-induced ISC by 37% in human IMCDi monolayers and fluid secretion by 91% in rat IMCDi. Immunohistochemistry of human medullary tissue sections revealed greater expression of {beta}2-AR than {beta}1-AR; {beta}2-AR was localized to the basolateral membranes of human IMCDi. Immunoblots identified {alpha}2A-AR and {alpha}2B-AR in cultured human IMCDi cell monolayers. We conclude that 1) catecholamines stimulate cAMPdependent anion and fluid secretion by IMCDi cells primarily through {beta}2-AR activation and 2) {alpha}2-AR activation attenuates cAMP-dependent anion secretion.




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