Tracing of labile zinc in live fish hepatocytes using FluoZin-3 |
| |
Authors: | Frederik A R Muylle Dirk Adriaensen Wim De Coen Jean-Pierre Timmermans Ronny Blust |
| |
Institution: | (1) Department of Biology, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp, Belgium;(2) Department of Biomedical Sciences, University of Antwerp, Groenenborgerlaan 171, B-2020 Antwerp, Belgium |
| |
Abstract: | Intracellular zinc levels are homeostatically regulated and although most is bound, a pool of labile Zn(II) is present in
cells. We show here that the zinc probe FluoZin-3 is useful to monitor zinc fluxes during fluorescent imaging of the trout
hepatic cell line D11. Nuclei and bulk cytosol appeared to lack detectable labile zinc, while the punctuate staining pattern
colocalized with a lysosome-specific probe. Applying extracellular zinc alone resulted in vesicular sequestration of the metal
ion. Together with Na-pyrithione a delayed and toxic rise in cellular fluorescence was triggered. When using another ionophore,
4-Br A23187, a zinc buffering effect of the vesicular pools was evident. Secondly, N-ethylmaleimide induced a homogeneous fluorescence rise, which was strongly enhanced by addition of Zn-pyrithione and disappeared
after TPEN washing. This suggests the involvement of thiol residues in controlling available cytosolic zinc. Our observations
have implications for the interpretation of calculated intracellular Zn2+ concentrations. |
| |
Keywords: | chelatable zinc fluorescent zinc probe confocal zinc imaging rainbow trout hepatocyte FluoZin-3 |
本文献已被 SpringerLink 等数据库收录! |
|