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Molecular and Functional Study of Ion Transporters in Membranes

Abstract

The group of Renal Biophysics of the Department of Physiology and Biophysics, ICB-USP, has in the past studied the mechanisms of ion transport in renal tubules of the mammalian kidney. Ion-sensitive microelectrodes for the determination of pH and potassium activity were develloped, including antimony and ion exchange resin based microelectrodes. These electrodes were used to study the mechanism of bicarbonate reabsorption and of potassium secretion in proximal and distal tubules of the renal cortex, as well as their regulation by hormonal factors. These studies benefited from the devellopment of a kinetic technique of analysis of stationary microperfusion experiments. More recently, the laboratory directed its interest toward more modern methods at the cellular and molecular level, including: 1- Molecular biology of membrane transporters, particularly of the Na/H exchanger and its isoforms, including the modulation of their expression during pathophysiological alterations of exchanger activity; 2- Determination of cellular ion activity by fluorescence microscopy, particularly of pH and calcium, in order to study mechanisms of cell homeostasis of these ions and their regulation; 3- Analysis of ion channels by the patch-clamp technique, including the properties of K and Ca channels in renal and other cells as well as the effect of spider toxins on these channels; 4- Functional characterization of Na-K dependent and of Na-dependent ATPases in insect renal tubules (Rhodnius, a transmitter of Chagas disease); and 5- Study of the mechanisms of volume regulation of insect and mammalian renal cells in hypotonic and hypertonic media. (AU)

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