Interference of heavy metal cations with fluorescent Ca2+ probes does not affect Ca2+ measurements in living cells

B. Marchi, B. Burlando, I. Panfoli, A. Viarengo

Risultato della ricerca: Contributo su rivistaArticolo in rivistapeer review

Abstract

In studies about the effects of heavy metals on intracellular Ca2+, the use of fluorescent probes is debated, as metal cations are known to affect the probe signal. In this study, spectrofluorimetric experiments in free solution, using Fluo-3 and Fura-2, showed that Zn2+ and Cd2+ enhanced the probe signal, Cu2+ quenched it, and Hg2+ had no effect. Addition of GSH prevented most of these effects, suggesting the occurrence of a similar protective role in living cells. Digital imaging of living mussel haemocytes loaded with Fura-2/AM or Fluo-3/AM showed that Hg2+, Cu2+ and Cd2+ induced a rise in probe fluorescence, whereas up to 200 μM Zn2+ had no effect. In particular, Cd2+ produced the strongest probe signal rise in free solution, but the lowest fluorescence increase in cells. Probe calibration yielded [Ca2+](i) values characteristic of resting levels in control and Zn2+-exposed cells, and, as expected, indicated Ca2+ homeostasis impairment in cells exposed to Cd2+, Cu2+ and Hg2+. Our results show that Ca2+ probe responses to heavy metals in living cells are completely different from those obtained in free solution, indicating that fluorescent probes can be a suitable tool to record the effects of heavy metals on [Ca2+](i). (C) 2000 Harcourt Publishers Ltd.

Lingua originaleInglese
pagine (da-a)225-231
Numero di pagine7
RivistaCell Calcium
Volume28
Numero di pubblicazione4
DOI
Stato di pubblicazionePubblicato - 2000
Pubblicato esternamente

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