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Pelzmann, B; Di Giuro, CM; Zorn-Pauly, K; Rossmann, C; Hallström, S; Groschner, K; Fameli, N.
Functional impairment of endothelial cells by the antimycotic amphotericin B.
Biochem Biophys Res Commun. 2016; 472(1):40-45
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Authors Med Uni Graz:
Di Giuro Cristiana Maria Loredana
Fameli Nicola
Groschner Klaus
Hallström Seth
Pelzmann Brigitte
Rossmann Christine Renate
Zorn-Pauly Klaus
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Abstract:
We set out to determine the membrane potential (Vm) of the endothelial cell line EA.hy926 and its sensitivity to the antimycotic amphotericin B (AmB), a commonly used antifungal component in cell culture media. We measured the endothelial Vm under various experimental conditions by patch clamp technique and found that Vm of AmB-treated cells is (-12.1 ± 9.3) mV, while in AmB-untreated (control) cells it is (-57.1 ± 4.1) mV. In AmB-free extracellular solutions, Vm recovered toward control levels and this gain in Vm rapidly dissipated upon re-addition of AmB, demonstrating a rapid and reversible effect of AmB on endothelial Vm. The consequences of AmB dependent alterations in endothelial transmembrane potential were tested at the levels of Ca(2+) signaling, of nucleotide concentrations, and energy metabolism. In AmB-treated cells we found substantially reduced Ca(2+) entry (to about 60% of that in control cells) in response to histamine induced endoplasmic reticulum (ER) Ca(2+) depletion, and diminished the ATP-to-ADP ratio (by >30%). Our data demonstrate a marked and experimentally relevant dependence of basic functional parameters of cultured endothelial cells on the presence of the ionophoric antimycotic AmB. The profound and reversible effects of the widely used culture media component AmB need careful consideration when interpreting experimental data obtained under respective culture conditions. Copyright © 2016 Elsevier Inc. All rights reserved.
Find related publications in this database (using NLM MeSH Indexing)
Adenosine Diphosphate - metabolism
Adenosine Triphosphate - metabolism
Amphotericin B - toxicity
Antifungal Agents - toxicity
Calcium Signaling - drug effects
Cell Culture Techniques - methods
Cell Line -
Culture Media - toxicity
Endothelial Cells - drug effects
Endothelial Cells - metabolism
Humans -
Membrane Potentials - drug effects
Patch-Clamp Techniques -

Find related publications in this database (Keywords)
Endothelium
EA.hy926
Membrane potential
Calcium signals
Depolarization
Metabolism
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