The Journal of General Physiology
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Published online 1 October 1999.
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© The Rockefeller University Press, 0022-1295/1999//561/ $5.00
Journal of General Physiology, Volume 114, Number 4, 1999


Original Article

Insulin Stimulates Transepithelial Sodium Transport by Activation of a Protein Phosphatase That Increases Na-K Atpase Activity in Endometrial Epithelial Cells

Chatsri Deachapunyaa, Melissa Palmer-Densmorea, and Scott M. O'Gradya

a From the Departments of Physiology and Animal Science, University of Minnesota, St. Paul, Minnesota 55108
Departments of Physiology and Animal Science, 495 Animal Science/Veterinary Medicine Building, University of Minnesota, St. Paul, Minnesota 55108.Fax: 612-625-2743;

ograd001{at}tc.umn.edu

The objective of this study was to investigate the effects of insulin and insulin-like growth factor I on transepithelial Na+ transport across porcine glandular endometrial epithelial cells grown in primary culture. Insulin and insulin-like growth factor I acutely stimulated Na+ transport two- to threefold by increasing Na+-K+ ATPase transport activity and basolateral membrane K+ conductance without increasing the apical membrane amiloride-sensitive Na+ conductance. Long-term exposure to insulin for 4 d resulted in enhanced Na+ absorption with a further increase in Na+-K+ ATPase transport activity and an increase in apical membrane amiloride-sensitive Na+ conductance. The effect of insulin on the Na+-K+ ATPase was the result of an increase in Vmax for extracellular K+ and intracellular Na+, and an increase in affinity of the pump for Na+. Immunohistochemical localization along with Western blot analysis of cultured porcine endometrial epithelial cells revealed the presence of {alpha}-1 and {alpha}-2 isoforms, but not the {alpha}-3 isoform of Na+-K+ ATPase, which did not change in the presence of insulin. Insulin-stimulated Na+ transport was inhibited by hydroxy-2-naphthalenylmethylphosphonic acid tris-acetoxymethyl ester [HNMPA-(AM)3], a specific inhibitor of insulin receptor tyrosine kinase activity, suggesting that the regulation of Na+ transport by insulin involves receptor autophosphorylation. Pretreatment with wortmannin, a specific inhibitor of phosphatidylinositol 3–kinase as well as okadaic acid and calyculin A, inhibitors of protein phosphatase activity, also blocked the insulin-stimulated increase in short circuit and pump currents, suggesting that activation of phosphatidylinositol 3–kinase and subsequent stimulation of a protein phosphatase mediates the action of insulin on Na+-K+ ATPase activation.

Key Words: insulin-like growth factor I • epithelial ion transport • membrane transport • ouabain • amiloride


© 1999 The Rockefeller University Press


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