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Re-evaluation of the expression of the gastric H,K-ATPase α subunit along the rat nephron
Authors:Lydie Cheval  Jean-Marc Elalouf  Alain Doucet
Affiliation:(1) Laboratoire de Biologie Intégrée des Cellules Rénales, Service de Biologie Cellulaire, CEA Saclay, Unité 1859 associée au CNRS, France, FR;(2) URA 1859-SBCe, Centre d’Etude de Saclay, F-91191 Gif sur Yvette cedex. France, FR
Abstract: Under normal conditions, the rat collecting duct displays an H,K-ATPase activity with kinetic and pharmacological properties very close to those of the gastric H,K-ATPase. However, whether the collecting duct H,K-ATPase and the gastric enzyme are identical remains controversial. Therefore, we re-evaluated the expression of the mRNAs encoding the gastric H,K-ATPase α subunit in the rat nephron. For this purpose, gastric H,K-ATPase mRNAs were quantitated by RT-PCR at the level of microdissected nephron segments using known amounts of gastric H,K-ATPase cRNA as external standards. Results indicate that gastric H,K-ATPase mRNAs are undetectable (<1 copy per cell) in the glomerulus and along the proximal tubule, thick ascending limb of Henle’s loop and collecting duct, although a faint expression (≈ 400 copies per μg total RNA) is measurable in whole–kidney preparations. Gastric H,K-ATPase mRNA is also absent along the nephron of K-depleted rats and of rats with chronic metabolic acidosis and alkalosis. Taken with other data from the literature, these results suggest that the collecting duct of normal rats might express an H,K-ATPase similar, but not identical, to the gastric isoform. Received: 29 October 1996 / Accepted: 21 November 1996
Keywords:  Gastric H  K-ATPase  Nephron segments  Collecting duct  RT-PCR  K depletion  Metabolic acidosis  Metabolic alkalosis
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