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November 1, 1978Proceedings of the National Academy of Sciences320 citationsOpen Access

Serum stimulates the Na+,K+ pump in quiescent fibroblasts by increasing Na+ entry.

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JSJeffrey B. SmithEREnrique Rozengurt

Key Points

  • To determine the mechanism by which serum stimulates Na+,K+ pump activity in quiescent 3T3 fibroblasts.
  • Measured 86Rb+ uptake and 22Na+ entry in quiescent 3T3 cells exposed to serum or sodium-carrying ionophores (monensin and gramicidin).
  • Assessed the sensitivity of the Na+,K+ pump to internal sodium concentrations and tested pump inhibition using ouabain in the presence or absence of external sodium.
  • Monensin and gramicidin markedly enhanced 86Rb+ uptake in a manner completely blocked by ouabain, showing that Na+,K+ pump activity is limited by and highly sensitive to internal Na+ levels.
  • Serum rapidly increased 22Na+ uptake and net Na+ entry during ouabain inhibition, while 0.3 µg/ml monensin replicated the exact magnitude of serum-induced pump activation.
  • Stimulation of 86Rb+ uptake by both serum and ionophores exhibited an absolute requirement for external Na+.

Abstract

Two ionophores (monensin and gramicidin) that carry Na+ into 3T3 cells markedly enhance the rate of 86Rb+ uptake. Ouabain prevents both ionophores from increasing 86Rb+ uptake, indicating that the ionophores activate the Na+,K+ pump. Measurements of 86Rb+ uptake and cell Na+ and K+ over a range of monensin concentrations show that the activity of the Na+,K+ pump in 3T3 cells is limited by the supply of internal Na+ and is extremely sensitive to small changes in internal Na+. Serum rapidly enhances the rate of 22Na+ uptake and net Na+ entry when Na+ exit is inhibited by ouabain. At 0.3 microgram/ml, monensin increases the rate of net Na+ entry and activates the Na+,K+ pump by the same degree as serum. The stimulation of 86Rb+ uptake by serum or the ionophores has an absolute requirement for external Na+. Thus, serum appears to stimulate the Na+,K+ pump in quiescent 3T3 cells by increasing its supply of Na+.

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Cite This Study

Smith et al. (1978) studied this question.

synapsesocial.com/papers/6a819cb233923b97f74cd513https://doi.org/10.1073/pnas.75.11.5560
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