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Quantitative radioautographic study of intracellular localization of calmodulin antagonist, W-7, in Chinese-hamster ovary cells by Y. Fujii; S. Ohno; H. Hidaka is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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The purpose of the present study was to analyse quantitatively the localization of calmodulin antagonist, n-(6-aminohexyl)-5-chloro-l-naphthalene-sulfonamide (W-7) in CHO-K1 cells. The cultured CHO-K1 cells were labelled with 1 (16.7 ~M), 2 (33.4 gM), 5 (83.5 gM) and 10 txCi/ml (167 gM) tritiated W-7. Some cells were preincubated in 10, 50 and 100 gM unlabelled W-7 for 30 min and then labelled with 2 or 5 gCi/ml tritiated W-7 for 1 h. The cells were doubly fixed in glutaraldehyde and osmium-tetroxide solution, and embedded in Epon. For light-microscopic radioautography, 2 gin-thick sections were wet mounted with radioautographic emulsion and exposed for 1 month. The radioautograms showed that large numbers of silver grains were mainly localized in the cytoplasm as well as in the nucleus. Quantitative analysis demonstrated that, in both the cytoplasm and nucleus, the number of silver grains was dependent on the concentration of the administered tritiated W-7 and the number was dramatically decreased by the pretreatment of unlabelled W-7. These results show that, in CHO-K1 cells, the W-7 binding sites are saturable. It is concluded that W-7 may get into CHO-KI cells and be bound to a

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Author

Y. Fujii; S. Ohno; H. Hidaka

Publisher

Springer; Springer-Verlag; Springer Verlag; Springer Science and Business Media LLC (ISSN 1432-119X)

Published

1985

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)