Comparative effect of ANF and various diuretics on isolated nephron segments

The effect of the synthetic form of ANF 0.1 to 10 microgram/ml (peptide 101-126), a diuretic and natriuretic peptide isolated from rat heart atria, on the metabolism of dog and rat kidney tubules was studied in vitro and compared to that of furosemide (0.1 to 1 mM), hydrochlorothiazide (0.5 mM) or amiloride (0.1 mM). In order to pinpoint eventual site(s) of ANF action along the nephron, proximal tubules, thick ascending limbs and papillary collecting ducts were isolated from dog kidneys as well as proximal tubules from rat kidneys. The substrate uptake (O2, lactate, glutamine, glucose) and production of metabolites (glutamate, ammonium, alanine, glucose) by these nephron segments were measured in absence or presence of the diuretic agents or the vehicle for ANF (acetate 1 mM). The total ATP turnover and the contribution of identified metabolic pathways for this turnover was calculated. It was expected that a molecule with diuretic properties reducing the permeability of cell membranes to NaCl would secondarily reduce the Na-K-ATPase activity, and therefore the oxygen and substrate utilization by affected cells. It was shown: that each nephron segment used presented the expected specific metabolic characteristics; that furosemide markedly inhibits the oxidative metabolism of thick ascending limbs; that acetate (the vehicle used for ANF) displaces the oxidation of glutamine and lactate in nephron segments with aerobic metabolism; that ANF had no effect on the metabolism of the studied segments despite the presence of specific c'GMP-generating receptors in the distal nephron. It is concluded that ANF must exert its natriuretic effect by a mechanism different from that of classical diuretics.