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Permanent link (DOI): https://doi.org/10.7939/R3MK65G1D

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The Role of Bicarbonate Transporters in the Development of Cardiovascular Diseases Open Access

Descriptions

Other title
Subject/Keyword
intracellular pH regulation
Bicarbonate transporters, carbonic anhydrase
Chloride/bicarbonate exchanger AE1
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Sowah, Daniel
Supervisor and department
Casey, Joseph (Biochemistry)
Examining committee member and department
Cole, William (University of Calgary)
Young, James (Physiology)
Fliegel, Larry (Biochemistry)
Light, Peter (Pharmacology)
Department
Department of Physiology
Specialization
Cell Biology
Date accepted
2014-01-31T13:37:40Z
Graduation date
2014-06
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
NHE1, whose activity and expression are elevated in cardiac hypertrophy, requires an acidifying pathway provided by AE3-mediated Cl-/HCO3- exchange to sustain its activity. Carbonic anhydrase II (CAII) interacts with NHE1 and AE3 to induce hypertrophy but the role of AE3 in hypertrophy is poorly elucidated. The present study seeks to characterize the interaction between CAII and AE1 as the prototype exchanger, and to delineate the role of AE3 in cardiac hypertrophy. AE1, the plasma membrane Cl-/HCO3- exchanger of erythrocytes, interacts with CAII to maximize HCO3− transport. To characterize further the effect of CAII on AE1 transport, CAII-wildtype or catalytically inactive CAII-V143Y was fused to the COOH terminus of AE1 to form AE1.CAII and AE1.CAII-V143Y, respectively. When expressed in HEK293 cells, AE1.CAII had a similar Cl-/HCO3- exchange activity to AE1 alone, as assessed by the flux of H+ equivalents (87 ± 4% vs. AE1) or rate of change of [Cl-]i (93 ± 4% vs. AE1). Contrastingly, AE1.CAII-V143Y displayed transport rates for H+ equivalents and Cl- of 55 ± 2% and of 40 ± 2%, versus AE1. AE1 fused to CAII-V143Y has reduced transport activity, which is compensated for during Cl-/HCO3− exchange by the presence of catalytically active CAII. When coexpressed with CAII-V143Y, AE1 bicarbonate transport was inhibited, whereas AE1.CAII activity was unaffected, suggesting impaired transport activity upon displacement of functional CAII from AE1 but not AE1.CAII. The role of AE3 in hypertrophy was studied in ae3 null mice (ae3-/-) whose disruption did not affect cardiac function as determined by echocardiography and blood pressure measurements. HW/BW ratio was larger in ae3+/+ compared to ae3-/- mice. Hypertrophic stimulation with phenylephrine or angiotensin II caused a 25-30% increase in surface area, upregulated ANP and β-BHC levels, and elevated protein synthesis of wildtype cardiomyocytes but ae3-/- cardiomyocytes were unaffected. Our findings confirm CAII association with AE1 increases Cl-/HCO3− exchange activity, consistent with the HCO3− transport metabolon model. Additionally, AE3 operating in conjunction with CAII and NHE1 is essential in neurohormonal-stimulated hypertrophy, forming a hypertrophic transport metabolon. Thus, antagonism of AE3 is an attractive target in the treatment of cardiac hypertrophy.
Language
English
DOI
doi:10.7939/R3MK65G1D
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication
Sowah, D., and Casey, J.R. (2011) An Intramolecular Transport Metabolon: Fusion of Carbonic Anhydrase II to the C-terminus of the Cl-/HCO3- Exchanger, AE1, Am. J. Physiol., 301, C336-46

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