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(Hypertension. 1998;31:1002-1006.)
© 1998 American Heart Association, Inc.

Scientific Contributions

Electroneutral Na-Coupled Cotransporter Expression in the Kidney During Variations of NaCl and Water Metabolism

Gabriela Moreno; Alejandra Merino; Adriana Mercado; Juan Pablo Herrera; Jorge González-Salazar; Ricardo Correa-Rotter; Steven C. Hebert; ; Gerardo Gamba

From the Molecular Physiology Unit, Department of Nephrology and Mineral Metabolism, Instituto Nacional de la Nutrición Salvador Zubirán and Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México (G.M., A. Merino, A. Mercado, J.P.H., J.G.-S., R.C.-R., G.G.); and the Division of Nephrology, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tenn (S.C.H.).

Correspondence to Gerardo Gamba, MD, PhD, Molecular Physiology Unit, Department of Nephrology and Mineral Metabolism, Instituto Nacional de la Nutrición Salvador Zubirán, Instituto de Investigaciones Biomédicas, UNAM, Vasco de Quiroga No. 15, Tlalpan 14000, Mexico City, México. E-mail gamba{at}mailer.main.conacyt.mx

Abstract—The purpose of the present study was to analyze the long-term regulation of renal bumetanide-sensitive Na⁺-K⁺-2Cl^- cotransporter and thiazide-sensitive Na⁺-Cl^- cotransporter gene expression during changes in NaCl and water metabolism. Male Wistar rats exposed to high or low NaCl intake, saline loading, dehydration, water loading, and furosemide administration during 7 days were studied. Control groups had access to regular food and tap water. Rats were kept in metabolic cages for 4 days before and during the experiment to determine daily urinary electrolyte excretion and osmolarity. At the end of the experiment, creatinine clearance and serum electrolyte levels were also measured. Kidneys were excised and macroscopically subdivided into cortex and outer and inner medulla. Total RNA was extracted from each individual cortex or outer medulla by use of the guanidine/cesium chloride method. The Na⁺-K⁺-2Cl^- cotransporter expression in outer medulla total RNA was assessed by nonradioactive Northern blot analysis and the Na⁺-Cl^- cotransporter expression in renal cortex total RNA was assessed by semiquantitative polymerase chain reaction. Experimental maneuvers were adequately tolerated, and all groups developed the appropriate renal response to each challenge. However, the level of expression of both cotransporters did not change in any model, except for a 2.8-fold increase in the Na⁺-Cl^- cotransporter expression during dehydration. We conclude that nephron adaptation to 7-day modifications in NaCl and water metabolism does not include changes in the amount of electroneutral sodium-coupled cotransporter gene expression at the mRNA level.

Key Words: bumetanide • thiazide • gene expression • ion transport

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