Na(+)-K(+)-Cl- cotransporter of brain capillary endothelial cells. Properties and regulation by endothelins, hyperosmolar solutions, calyculin A, and interleukin-1.
Cultured rat brain capillary endothelial cells expressed a large 86Rb+ uptake component that was dependent on external Na+ and Cl- and that was inhibited by loop diuretics with unusual pharmacological properties: benzmetanide (IC50 = 1-5 microM) = bumetanide (IC50 = 1-5 microM) > piretanide (IC50 = 3-16 microM) = furosemide (IC50 = 7-11 microM). It was activated 2-fold by endothelin-1 (EC50 = 1 nM) and endothelin-3 (EC50 = 9 nM). The actions of endothelins were prevented by BQ-123 (cyclo-(D-Trp-D-Asp-Pro-D-Val-Leu)) in a competitive manner and with a high affinity, thus indicating the involvement of an atypical BQ-123-sensitive, ETA-like receptor that had a high affinity for endothelin-3. Neither protein kinase C nor Ca(2+)-dependent protein kinases mediated the actions of endothelins. Cotransport activity was increased 4-fold by hyperosmotic cell shrinkage. Basal Na(+)-K(+)-Cl- cotransport activity was partially inhibited by isoproterenol and was unaffected by agents that promoted cGMP formation. Calyculin A, an inhibitor of protein phosphatases, stimulated cotransport activity and potentiated the action of endothelin-1, but not that of cell shrinkage. Basal and stimulated cotransport activities were inhibited by genistein, a protein kinase inhibitor with similar potencies, and by staurosporine, which has different potencies. Finally, endothelin-1-stimulated activity was partially and specifically inhibited by interleukin-1. It is concluded that rat brain capillary endothelial cells express a Na(+)-K(+)-Cl- cotransporter that has unique properties and that is regulated by multiple protein kinase/phosphatase systems. It is a target for low concentrations of endothelins and may play a role in brain-to-blood movements of K+.
Vigne
P
P
Institut de Pharmacologie Moléculaire et Cellulaire, CNRS, Université de Nice-Sophia Antipolis, Valbonne, France.
Lopez Farre
A
A
Frelin
C
C
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D001921:Brain / Q000098:blood supply / Q000378:metabolism*; D002196:Capillaries / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D017869:Cyclic GMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D016232:Endothelins / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000378:metabolism*; D007375:Interleukin-1 / Q000494:pharmacology*; D008387:Marine Toxins; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D010080:Oxazoles / Q000494:pharmacology*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D051381:Rats; D015398:Signal Transduction; D028021:Sodium-Potassium-Chloride Symporters
D002352:Carrier Proteins; D016232:Endothelins; D007375:Interleukin-1; D008387:Marine Toxins; D008565:Membrane Proteins; D010080:Oxazoles; D028021:Sodium-Potassium-Chloride Symporters; C059041:calyculin A; D017868:Cyclic AMP-Dependent Protein Kinases; D017869:Cyclic GMP-Dependent Protein Kinases; D010749:Phosphoprotein Phosphatases