Ouabain uptake by endocytosis in isolated guinea pig atria.
Mammalian cells specifically internalize some molecular species through receptor-mediated endocytosis (RME). We have used four different experimental protocols to investigate whether ouabain enters cardiac cells of guinea pig atrium through this pathway. First, by electron microscope morphometry we found that ouabain increased endocytic vesicles in atrial cells. Second, by scintillation counting we found that [3H]ouabain uptake by the tissue is decreased by three treatments that decrease RME, i.e., NH4Cl, trifluoperazine, and 16 mM [K+]0. Third, by radioautography at the electron microscope level, we checked that in preceding experiments [3H]ouabain was washed out of plasma membrane after 60-min rinse and interiorized into the cardiac cells. Fourth, isometric tension recordings showed that the positive inotropic effect of ouabain was diminished in the presence of inhibitors, whereas that of a hydrophobic analogue, ouabagenin, was not affected. These results suggest that ouabain enters cardiac cells through RME and also that an intracellular site may, at least in part, be responsible for its inotropic effect.
Núñez-Durán
H
H
Departamento de Ciencias Fisiológicas, Universidad Autónoma de Puebla, Mexico.
Riboni
L
L
Ubaldo
E
E
Kabela
E
E
Bárcenas-Ruiz
L
L
D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport; D004705:Endocytosis; D006168:Guinea Pigs; D006325:Heart Atria / Q000378:metabolism / Q000648:ultrastructure; D066298:In Vitro Techniques; D007700:Kinetics; D008854:Microscopy, Electron; D009200:Myocardial Contraction / Q000187:drug effects*; D009206:Myocardium / Q000378:metabolism* / Q000648:ultrastructure; D010042:Ouabain / Q000031:analogs & derivatives / Q000378:metabolism* / Q000494:pharmacology; D011955:Receptors, Drug / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase; D014268:Trifluoperazine / Q000494:pharmacology
D011955:Receptors, Drug; C028996:cardiac glycoside receptors; D000643:Ammonium Chloride; D014268:Trifluoperazine; D010042:Ouabain; D000254:Sodium-Potassium-Exchanging ATPase; C008721:ouabagenin