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Am J Physiol Renal Physiol (September 13, 2005). doi:10.1152/ajprenal.00395.2004
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Submitted on October 29, 2004
Accepted on August 31, 2005

PMA- and ANGII-induced PKC regulation of the renal Na+/HCO3- cotransporter (hkNBCe1)

Clint Perry1, Judith Blaine1, Hong Le1, and Irina I. Grichtchenko1*

1 Department of Physiology and Biophysics, UCHSC, Aurora, CO, USA

* To whom correspondence should be addressed. E-mail: irina.grichtchenko{at}uchsc.edu.

The renal electrogenic Na/HCO3 - cotransporter (hkNBCe1) plays a major role in the bicarbonate reabsorption by the kidney. We examined how PMA- and ANGII-activated PKCs regulate hkNBCe1 expressed with or without the ANGII receptors AT1B in Xenopus oocytes. We found that 10 nM PMA halved the hkNBCe1 current detected in voltage-clamped oocytes. A PKC-specific inhibitor GF109203X, and a specific inhibitor of Ca-dependent conventional PKC{alpha}{beta}{gamma}, GO6976, significantly reduced PMA inhibition. PMA did not alter surface expression of the cotransporters, but it significantly increased hkNBCe1-PKC{alpha}{beta}{gamma} membrane association. We found that at 10-6 M, ANGII halved the hkNBCe1 current detected in oocytes co-expressing cotransporters with AT1B. A PKC-specific inhibitor GF109203X, and a PKC{epsilon} translocation inhibitor {epsilon}V1-2 peptide as well as BAPTA-AM (but not GO6976) significantly reduced ANGII inhibition. At 10-6 M, ANGII significantly decreased surface expression of the cotransporters and increased hkNBCe1-PKC{epsilon} membrane association. Additionally, we found that at 10-11 and 10-10 M ANGII stimulated hkNBCe1 current. This effect was blocked by BAPTA-AM and partially reduced by GF109203X. We also found that ANGII increased intracellular Ca2+ in fluo-4-loaded oocytes. Our results suggest that: (I) PMA-inhibition of hkNBCe1 is mediated by Ca-dependent PKC{alpha}{beta}{gamma} and 10nM PMA does not induce downregulation of cotransporter surface expression. (II) 10-6 M ANGII-inhibition of hkNBCe1 is mediated by both Ca-independent PKC{epsilon} and downregulation of cotransporter surface expression, possibly triggered by intracellular Ca2+ mobilization. (III) Similar to proximal tubule, acute ANGII has a biphasic effect on hkNBCe1 co-expressed with AT1B In Xenopus oocytes.




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