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We used patch-clamp and Western blot analysis to test whether PGF 2α  stimulates the basolateral 10-pS Cl −  channel and thiazide-sensitive Na + -Cl −  cotransporter (NCC) in the distal convoluted tubule (DCT) via a prostaglandin F receptor (FP-R). Single channel and whole cell recordings demonstrated that PGF 2α  stimulated the 10-pS Cl −  channel in the DCT. The stimulatory effect of PGF 2α  on the Cl −  channel was mimicked by a FP-R agonist, latanoprost, but was abrogated by blocking FP-R with AL8810. Also, the effect of PGF 2α  on the Cl −  channel in the DCT was recapitulated by stimulating PKC but was blocked by inhibiting PKC. Furthermore, inhibition of p38 MAPK but not ERK blocked the effect of PGF 2α  on the 10-pS Cl −  channel. Inhibition of NADPH oxidase also abrogated the stimulatory effect of PGF 2α  on the 10-pS Cl −  channel, while the addition of 10 μM H 2 O 2  mimicked the stimulatory effect of PGF 2α  on the 10-pS Cl −  channel. Moreover, superoxide-related species may mediate the stimulatory effect of PGF 2α  on the 10-pS Cl −  channel because the stimulatory effect of PGF 2α  and H 2 O 2  was not additive. Western blot analysis showed that infusion of PGF 2α  in vivo not only increased the expression of FP-R but also increased the expression of total NCC and phosphorylated NCC. We conclude that PGF 2α  stimulates the basolateral 10-pS Cl −  channel in the DCT by activating FP-R through PKC/p38 MAPK and NADPH oxidase-dependent pathways. The stimulatory effects of PGF 2α  on the Cl −  channel and NCC may contribute to PGF 2α -induced increases in NaCl reabsorption in the DCT.

american journal of physiology. renal physiology./american journal of physiology. renal physiology

PGF2α stimulates the 10-pS Cl− channel and thiazide-sensitive Na+-Cl− cotransporter in the distal convoluted tubule