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
AbstractThe 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.
© 1998 American Heart Association, Inc.
Scientific Contributions
Electroneutral Na-Coupled Cotransporter Expression in the Kidney During Variations of NaCl and Water Metabolism
Key Words: bumetanide thiazide gene expression ion transport
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