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1 Department of Physiology and Biophysics, University of South Florida, Tampa, Florida 33612; and 2 Department of Molecular Pharmacology, Physiology and Biotechnology, Brown University, Providence, Rhode Island 02912
We previously showed that acute arterial hypertension induces an inhibition of fluid and NaCl reabsorption in proximal tubules of Sprague-Dawley rats, which is associated with a rapid reversible internalization of apical Na+/H+ exchanger in brush border. To determine whether there is a corresponding inhibition of apical Na+/H+ exchanger activity in proximal tubules to account for the reduced tubular reabsorption, an instrument capable of measuring intracellular pH (pHi) ratiometrically and repeatedly on the surface of kidney with high temporal resolution is required. We report the design and validation of such a fluorimetric system based on two ultraviolet nitrogen-pulsed lasers and a photomultiplier. pHi of proximal tubules in situ was measured with pH-sensitive fluorescence dye 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein at 5 Hz. Using the initial rate of change of pHi (dpHi/dt) during luminal Na+ removal as an index of apical Na+/H+ exchanger activity, the exchanger activity was found to be reduced by 52 ± 11% (n = 14, P < 0.05) compared with the baseline after 20 min of induced acute hypertension. The inhibition of Na+/H+ exchange activity was alleviated when the blood pressure was returned to prehypertensive level. These observations indicate that acute changes in arterial pressure can reversibly inhibit apical Na+/H+ exchanger activity, which might contribute to pressure natriuresis in proximal tubule.
intracellular pH, 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein, pressure natriuresis, microperfusion
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