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J. Biol. Chem., Vol. 266, Issue 5, 2783-2788, Feb, 1991
L Hunyady, F Merelli, AJ Baukal, T Balla and KJ Catt
The relationships between receptor-mediated endocytosis and the generation
of intracellular signals were analyzed in angiotensin II (AII)-stimulated
adrenal glomerulosa cells. In cells equilibrated with 125I-AII analogs at 4
degrees C, specifically bound agonist but not antagonist AII derivatives
were rapidly internalized at 37 degrees C. AII-induced internalization was
not influenced by the presence or absence of extracellular Ca2+ but was
inhibited by treatment with phenylarsine oxide (PAO) or by arresting coated
pit formation with hypotonic shock and potassium depletion. Inhibition of
internalization by PAO was prevented by the bifunctional sulfhydryl reagent
dithiothreitol but only partially reversed by mercaptoethanol, and
readdition of K+ restored internalization in K(+)-depleted cells. Treatment
with PAO did not impair the initial AII-induced elevations of inositol
1,4,5-trisphosphate (Ins(1,4,5)P3) and cytoplasmic calcium [( Ca2+]i) but
reduced the sustained phase of the Ins(1,4,5)P3 response by 85% and
abolished the second phase of the cytoplasmic Ca2+ response; these
responses were restored by concomitant treatment with dithiothreitol.
Inhibition of AII-receptor internalization by K+ depletion also caused
selective loss of the sustained phase of the AII- induced Ca2+ response.
Thus, blockade of AII-receptor internalization has similar effects as
extracellular Ca2+ deficiency, which abolishes the sustained but not the
early AII-induced increases in Ins(1,4,5)P3 production and [Ca2+]i. The
close correlations between AII-induced internalization and the generation
of Ins(1,4,5)P3 and [Ca2+]i responses suggest that endocytosis of the
agonist-receptor complex is necessary to maintain the production of these
intracellular signals. It is also possible that receptor-operated vesicular
uptake of extracellular Ca2+ makes a significant contribution to the
sustained [Ca2+]i responses of certain agonist-stimulated target cells.
Agonist-induced endocytosis and signal generation in adrenal glomerulosa cells. A potential mechanism for receptor-operated calcium entry
Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
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