NDLI: Validation of TPEN as a Zinc Chelator in Fluorescence Probing of Calcium in Cells with the Indicator Fura-2

Content Provider	Springer Nature Link
Author	Matias, Carlos M. Sousa, João M. Quinta Ferreira, M. Emília Arif, Mona Burrows, Hugh D.
Copyright Year	2009
Abstract	Fura-2 is widely used as a fluorescent probe to monitor dynamic changes in cytosolic free calcium in cells, where Ca$^{2+}$ can enter through several types of voltage-operated or ligand-gated channels. However, Fura-2 is also sensitive to other metal ions, such as zinc, which may be involved in ionic channels and receptors. There is interest, in particular, in studying the synapses between mossy fibers and CA3 pyramidal cells which contain both calcium and high quantities of free or loosely bound zinc. We have found, through fluorescence probing, that endogenous zinc inhibits mossy fiber calcium transients. However, since these results might be explained by an effect of the zinc chelator N,N,N’,N’-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN) on the spectral properties of Fura-2, we have carried out a validation of the method through fluorescence excitation spectra of the complex Fura-2/calcium, and show that TPEN does not affect these spectra. This supports the idea that the observed calcium enhancement is related to a zinc inhibition of presynaptic calcium mechanisms, and confirms the use of the chelator TPEN as a general procedure for the biophysical study of Ca(II) in the presence of Zn(II) using Fura-2.
Starting Page	377
Ending Page	380
Page Count	4
File Format	PDF
ISSN	10530509
Journal	Journal of Fluorescence
Volume Number	20
Issue Number	1
e-ISSN	15734994
Language	English
Publisher	Springer US
Publisher Date	2009-10-10
Publisher Place	Boston
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Fluorescence probing Cellular calcium sensing TPEN zinc chelator Fura-2 Analytical Chemistry Biochemistry Biotechnology Biophysics and Biological Physics Biomedicine general
Content Type	Text
Resource Type	Article
Subject	Sociology and Political Science Spectroscopy Law Biochemistry Clinical Biochemistry Clinical Psychology

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