Receptor-assay for endogenous inhibitors of Na-K ATPase

It is now accepted that there is an endogenous digitalis-like substance (EDLS), previously called natriuretic factor, present in different mammalian species which participates to the regulation of the sodium balance and at least in some situations, to the genesis of hypertension. The physiological and pathophysiological role of this substance is well recognized. However, its chemical nature remains elusive. Our purpose was to define a receptor assay for this EDLS in order to be able to measure its concentration in urine and blood. To do so, we first prepared a pool of lyophylised urine from salt loaded men. The active material was isolated by Sephadex G25 chromatography. Using the same active fraction, we investigated its biological effects. Series of experiments have demonstrated the various properties of the EDLS: inducing natriuresis when injected into a rat, diminishing short circuit current in toad's bladder and colonic mucosa of rat in in vitro preparations, inhibiting Na-K ATPase activity in isolated toad's bladder cells, binding to ouabain receptors and cross-reacting with different antibodies directed against digoxin. Among these properties, the binding to ouabain receptors offers the unique opportunity to progress in our knowledges of the substance, making possible the calculation of its constant of affinity and the estimation of the molality of the active material contained in a gram of crude extract. Knowing the constant of affinity of the EDLS for its receptor would provide the best identification of the substance as long as we do not know its exact chemical nature.(ABSTRACT TRUNCATED AT 250 WORDS)