Stimulation of na+-k+-2cl- cotransport in rat medullary thick ascending limb by dopamine. Aoki, Yuji, Frederick E. Albrecht, Kenneth R. Bergman, and Pedro A. Jose. The Second Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto, Japan, Division of Pediatric Nephrology, Georgetown University Children's Medical Center, and Department of Physiology and Biophysics, Georgetown University School of Medicine, Washington, DC 20007
APStracts 3:0204R, 1996.
Dopamine receptors are present in the medullary thick ascending limb (mTAL) of Henle but their effect on ion transport in this nephron segment has not been tested. Therefore, we studied the short-term effects of dopamine on Na+-K+-2Cl- cotransport (assessed by 100 [mu]M bumetanide-sensitive 86Rb uptake) in rat mTAL tubular suspensions. Dopamine (1 [mu]M ) stimulated bumetanide-sensitive 86Rb uptake (72.1+/-10.6% vs control, n=5) by increasing total 86Rb uptake and by decreasing bumetanide-insensitive 86Rb ptake; this effect was concentration-dependent. The dopamine-induced stimulation of Na+-K+-2Cl- cotransport activity was mimicked by calyculin A, a protein phosphatase (PP) inhibitor, and Sp-cAMPS, a protein kinase A (PKA) agonist and blocked by Rp-8-CPT-cAMPS, a PKA inhibitor (n=5). Dopamine did not increase the stimulatory effect of the PP inhibitor. However, the stimulatory effect of the PP inhibitor and PKA agonist was additive and approached the stimulatory effect of dopamine. The stimulatory effects of dopamine, PP inhibitor and PKA agonist persisted even when intracellular sodium was clamped by 5 [mu]M monensin. When K+ channels were blocked by 1 mM BaCl2, the effects of dopamine and calyculin A on the cotransport were no longer apparent while the stimulatory effect of the PKA agonist was attenuated. We conclude that dopamine stimulates Na+-K+-2Cl- cotransport activity. This action is mediated mainly by PKA-dependent phosphorylation/dephos-phorylation processes and modulated by dopamine actions on K+ channels.

Received 19 January 1996; accepted in final form 28 May 1996.
APS Manuscript Number R29-6.
Article publication pending Am. J. Physiol. (Regulatory Integrative
Comp. Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 17 June 96