Participation of protein phosphatases in regulation of sodium transport across the erythrocyte membrane of the lamprey Lampetra fluviatilis

Abstract

Erythrocytes of lamprey Lampetra fluviatilis were incubated in standard isotonic medium at 20°C with 22Na to determine the unidirectional Na+ influx. Cell incubation in the presence of various protein phosphatase inhibitors (NaF, cantharidin, calyculin A) led to a considerable increase of Na+ transport into erythrocytes. The stimulation of Na+ influx into erythrocytes rose with increase of concentration of calyculin A within the range of 10–100 nM. The calyculin A concentration producing a 50% activation of Na+ transport amounted to 41.5 nM. Under optimal experimental conditions, the Na+ influx increased from control level of 5–8 to 20–40 mmol/l cells/h under effect of protein phosphatase blockers. The Na+ transport induced by these inhibitors was completely suppressed on addition of amiloride to the incubation medium. The treatment of lamprey erythrocytes with protein phosphatase inhibitors was accompanied by a small (∼12%), but statistically significant decrease of intracellular Na+ content. A small decrease of intracellular K+ content in erythrocyte was observed only under the effect of NaF. The obtained data allow making the conclusion that protein phosphatases of the PP1 and PP2A types play a significant role in regulation of Na+ transport across the lamprey erythrocyte membrane in both directions.