Abstract 116: Characterization of a Sodium Responsive Human Sodium Bicarbonate Transporter NBCe2 in Human Proximal Tubule
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Abstract
The electrogenic sodium bicarbonate cotransporter (NBCe2) is encoded by SLC4A5, variants of which have been associated with salt sensitivity of blood pressure (BP), which affects 25% of the adult population. NBCe2 is thought to mediate sodium bicarbonate cotransport primarily in the renal collecting duct, but NBCe2 mRNA is also found in the rodent renal proximal tubule (RPT), another site of bicarbonate transport. The protein expression or function of NBCe2 has not been demonstrated in the human RPT. We validated an NBCe2 antibody by siRNA and Western blot, as well as overexpression of an epitope-tagged NBCe2 construct in both RPT cells (RPTCs) and HEK293 cells. We immune-localized (peptide pre-absorbable) NBCe2 protein in the RPT of fresh and frozen human kidney slices, RPTCs isolated from human urine, and in isolated RPTC apical membrane. NBCe2 was primarily found in the Golgi while NBCe1, another electrogenic member of the same family, was primarily found at the basolateral membrane, under basal conditions. Following a short-term increase in intracellular sodium, NBCe2 expression increased at the apical membrane in cultured slices of human kidney (1.75±0.15 fold increase over basal, N=3, p<0.05) and polarized, immortalized RPTCs (0.08±0.015 vs 0.03±0.006 control, NBCe2/CD13 RFU, N=3, p<0.05), while the basolateral localization of NBCe1 was decreased (30.7±5.4% over basal, N=3, p<0.05). DIDS (electrogenic sodium bicarbonate inhibitor, 500 μM, 10 min) inhibitable bicarbonate dependent pH recovery in the presence of EIPA (NHE inhibitor,100 nM, 10 min) after ammonium chloride prepulse (20 mM, 5 min) was higher in the cells cultured under high salt conditions (26.7±7.2% over basal, N=3, p<0.05). Thus, NBCe2 appears to be important in apical sodium and bicarbonate cotransport under high salt conditions. Future studies will examine the role of NBCe2 in mediating increased renal sodium transport in human salt sensitivity of BP.
Author Disclosures: J.J. Gildea: A. Employment; Significant; The University of Virginia. B. Research Grant (includes principal investigator, collaborator, or consultant and pending grants as well as grants already received); Significant; NHLBI. P. Xu: A. Employment; Significant; University of Virginia. M. Carlson: None. R.T. Gaglione: None. D. Bigler Wang: None. B.A. Kemp: A. Employment; Significant; The University of Virginia. C.M. Reyes: None. H.E. McGrath: A. Employment; Significant; The University of Virginia. R.M. Carey: A. Employment; Significant; The University of Virginia. B. Research Grant (includes principal investigator, collaborator, or consultant and pending grants as well as grants already received); Significant; NHLBI. P.A. Jose: A. Employment; Significant; The University of Maryland. B. Research Grant (includes principal investigator, collaborator, or consultant and pending grants as well as grants already received); Significant; NHLBI. R.A. Felder: A. Employment; Significant; The University of Virginia. B. Research Grant (includes principal investigator, collaborator, or consultant and pending grants as well as grants already received); Significant; NHLBI.
- © 2015 by American Heart Association, Inc.
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- Abstract 116: Characterization of a Sodium Responsive Human Sodium Bicarbonate Transporter NBCe2 in Human Proximal TubuleJohn J Gildea, Peng Xu, M Carlson, Robert T Gaglione, Dora Bigler Wang, Brandon A Kemp, Camellia M Reyes, Helen E McGrath, Robert M Carey, Pedro A Jose and Robin A FelderHypertension. 2015;66:A116, originally published November 3, 2015
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- Abstract 116: Characterization of a Sodium Responsive Human Sodium Bicarbonate Transporter NBCe2 in Human Proximal TubuleJohn J Gildea, Peng Xu, M Carlson, Robert T Gaglione, Dora Bigler Wang, Brandon A Kemp, Camellia M Reyes, Helen E McGrath, Robert M Carey, Pedro A Jose and Robin A FelderHypertension. 2015;66:A116, originally published November 3, 2015