Real-time assessment of α-ketoglutarate effect on organic anion secretion in perfused rabbit proximal tubules

Abstract

To determine the quantitative roles of the basolateral and luminal Na+-dicarboxylate (Na-DC) cotransporters in establishing and maintaining the α-ketoglutarate (αKG) gradient required for renal tubular secretion of organic anions, we measured net steady-state transepithelial secretion of fluorescein (FL) in real time in isolated, perfused S2 segments of rabbit renal proximal tubules. Net “basal” FL secretion in the absence of exogenous αKG had a K t of ∼4 μM and a maximal transepithelial secretion rate ( J max) of ∼380 fmol ⋅ min−1 ⋅ mm−1(where K t is the FL concentration that produces one-half the J max). It could be almost completely inhibited by basolateral p-aminohippurate (PAH). Selective inhibition of the basolateral Na-DC cotransporter indicated that recycling via this transporter of αKG that had been exchanged for FL supports ∼25% of the “basal” FL secretion. Physiological αKG concentrations of 10 μM in the bath or 50 μM in the perfusate stimulated net secretion of FL by ∼30 or ∼20%, respectively. These data indicate that the basolateral Na-DC cotransporter supports ∼42% of the net FL secretion. The luminal and basolateral effects of physiological concentrations of αKG were additive, indicating that the combined function of the luminal and basolateral Na-DC cotransporters can support ∼50% of the net FL secretion. This apparently occurs by their establishing and maintaining ∼50% of the outwardly directed αKG gradient that is responsible for driving basolateral FL/αKG exchange. The remaining ∼50% would be maintained by metabolic production of αKG in the cells.