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Am J Physiol Cell Physiol 255: C479-C485, 1988;
0363-6143/88 $5.00
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AJP - Cell Physiology, Vol 255, Issue 4 C479-C485, Copyright © 1988 by American Physiological Society

ARTICLES

Ouabain uptake by endocytosis in isolated guinea pig atria

H. Nunez-Duran, L. Riboni, E. Ubaldo, E. Kabela and L. Barcenas-Ruiz
Departamento de Ciencias Fisiologicas, Universidad Autonoma de Puebla, Mexico.

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.


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