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Am J Physiol Renal Physiol 274: F165-F174, 1998;
0363-6127/98 $5.00
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Vol. 274, Issue 1, F165-F174, January 1998

Effect of alpha -ketoglutarate on organic anion transport in single rabbit renal proximal tubules

John R. Welborn, Shlomo Shpun, William H. Dantzler, and Stephen H. Wright

Department of Physiology, College of Medicine, University of Arizona, Tucson, Arizona 85724

The effect of exogenous alpha -ketoglutarate (alpha KG) and the peritubular Na+-dicarboxylate (Na-DC) cotransporter on organic anion/dicarboxylate (OA/DC) exchange in S2 segments of single, nonperfused rabbit proximal tubules was measured using 1 µM fluorescein (FL), a model OA, and epifluorescence microscopy. The effect of different transmembrane distributions of 10 µM alpha KG on peritubular FL uptake was measured at 37°C using bicarbonate-buffered, nutrient-containing buffers, which are conditions similar to those found in vivo. Compared with FL uptake in the absence of exogenous alpha KG, preloading tubules with alpha KG (trans-configuration) or acute exposure to alpha KG (cis-configuration) increased FL uptake 62% and 54%, respectively, whereas a cis-trans-configuration of alpha KG increased FL uptake by 76%. The cis-stimulation of FL uptake by alpha KG was rapid, within 5-7 s. This stimulation was blocked 96% by simultaneous exposure to 2 mM Li+, indicating that stimulation of transport was secondary to the uptake of exogenous alpha KG. In the absence of exogenous alpha KG, selective inhibition of Na-DC cotransport using 2 mM Li+ or 1 mM methylsuccinate decreased FL uptake by 25% (effects that were reversible but not additive), suggesting that the Na-DC cotransporter recycles endogenous alpha KG that has left the cell in exchange for FL and that this activity supports ~25% of baseline activity of the OA/DC exchanger. With recycling of alpha KG accounting for ~25% of FL uptake and with accumulation of exogenous alpha KG accounting for another ~75% increase in FL uptake, Na-DC cotransport appears to directly support (25% + 75%)/175%, or ~57%, of total FL transport.

kidney; metabolic intermediates; fluorescein; cotransport; epifluorescence microscopy


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