title: Regulation of K-Cl cotransport in erythrocytes of frog Rana temporaria by commonly used protein kinase and protein phosphatase inhibitors.
abstract: Recently (Agalakova and Gusev in J Comp Physiol 179:443-450, 2009), we demonstrated that the activity of K-Cl cotransport (KCC) in frog red blood cells is inhibited under stimulation of protein kinase C (PKC) with phorbol ester PMA (12-myristate-13-acetate). Present work was performed to uncover possible implication of protein kinases and protein phosphatases (PPs) in the regulation of baseline and volume-dependent KCC activity in these cells. K+ influx was estimated as 86Rb uptake by the cells in isotonic or hypotonic media in the presence of ouabain, K+ efflux was determined as the difference between K+ loss by the cells incubated in parallel in isotonic or hypotonic K(+)-free Cl(-)- and NO(3)(-)-media. Swelling of the cells in hypotonic medium was accompanied by approximately 50% activation of Cl-dependent K+ influx and efflux. Protein tyrosine kinase (PTK) inhibitor genistein (0.1 mM) stably and considerably (up to 89%) suppressed both baseline and volume-dependent KCC activity in each direction. Other PTK blockers (tyrphostin 23 and quercetin) had no influence on KCC activity in frog erythrocytes. PKC inhibitor chelerythrine (20 microM) and both PP inhibitors, fluoride (5 mM) and okadaic acid (1 microM), reduced KCC activity by 25-70%. Neither basal nor swelling-activated KCC in frog erythrocytes was affected by PKC inhibitor staurosporine (1 microM). Based on the previous and present results, we can suggest that the main role in the maintenance of basal and volume-dependent KCC activity in frog erythrocytes belongs to PTKs and PPs, whereas PKC is a negative regulator of this ion system.
authors: Sechenov Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, Thorez pr. 44, 194223 St. Petersburg, Russia.
authors: Gennadii Petrovich
authors: GP
authors: Gusev
authors: Natalia Ivanovna
authors: NI
authors: Agalakova
mesh_terms: D000818:Animals; D053119:Benzophenanthridines / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism*; D005459:Fluorides / Q000494:pharmacology; D019833:Genistein / Q000494:pharmacology; D019319:Okadaic Acid / Q000494:pharmacology; D010042:Ouabain / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000502:physiology*; D011188:Potassium / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D011794:Quercetin / Q000494:pharmacology; D011896:Rana temporaria; D019311:Staurosporine / Q000494:pharmacology; D027981:Symporters / Q000187:drug effects / Q000502:physiology*; D020032:Tyrphostins / Q000494:pharmacology; D000096922:K Cl- Cotransporters
chemical_list: D053119:Benzophenanthridines; D004791:Enzyme Inhibitors; D005459:Fluorides; D019833:Genistein; D019319:Okadaic Acid; D010042:Ouabain; D010749:Phosphoprotein Phosphatases; D011188:Potassium; D011493:Protein Kinase C; D047428:Protein Kinase Inhibitors; D011505:Protein-Tyrosine Kinases; D011794:Quercetin; D019311:Staurosporine; D027981:Symporters; D020032:Tyrphostins; D000096922:K Cl- Cotransporters; C016299:chelerythrine; C060641:tyrphostin A23