Papers

Peer-reviewed
May, 2015

Potential Interactions of Calcium-Sensitive Reagents with Zinc Ion in Different Cultured Cells

PLOS ONE
  • Koichi Fujikawa
  • ,
  • Ryo Fukumori
  • ,
  • Saki Nakamura
  • ,
  • Takaya Kutsukake
  • ,
  • Takeshi Takarada
  • ,
  • Yukio Yoneda

Volume
10
Number
5
First page
e0127421
Last page
Language
English
Publishing type
Research paper (scientific journal)
DOI
10.1371/journal.pone.0127421
Publisher
PUBLIC LIBRARY SCIENCE

Background
Several chemicals have been widely used to evaluate the involvement of free Ca2+ in mechanisms underlying a variety of biological responses for decades. Here, we report high reactivity to zinc of well-known Ca2+-sensitive reagents in diverse cultured cells.
Methodology/Principal Findings
In rat astrocytic C6 glioma cells loaded with the fluorescent Ca2+ dye Fluo-3, the addition of ZnCl2 gradually increased the fluorescence intensity in a manner sensitive to the Ca2+ chelator EGTA irrespective of added CaCl2. The addition of the Ca2+ ionophore A23187 drastically increased Fluo-3 fluorescence in the absence of ZnCl2, while the addition of the Zn2+ ionophore pyrithione rapidly and additionally increased the fluorescence in the presence of ZnCl2, but not in its absence. In cells loaded with the zinc dye FluoZin-3 along with Fluo-3, a similarly gradual increase was seen in the fluorescence of Fluo-3, but not of FluoZin-3, in the presence of both CaCl2 and ZnCl2. Further addition of pyrithione drastically increased the fluorescence intensity of both dyes, while the addition of the Zn2+ chelator N, N, N', N'-tetrakis(2-pyridylmethyl) ethane-1,2-diamine (TPEN) rapidly and drastically decreased FluoZin-3 fluorescence. In cells loaded with FluoZin-3 alone, the addition of ZnCl2 induced a gradual increase in the fluorescence in a fashion independent of added CaCl2 but sensitive to EGTA. Significant inhibition was found in the vitality to reduce 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide in a manner sensitive to TPEN, EDTA and BAPTA in C6 glioma cells exposed to ZnCl2, with pyrithione accelerating the inhibition. Similar inhibition occurred in an EGTA-sensitive fashion after brief exposure to ZnCl2 in pluripotent P19 cells, neuronal Neuro2A cells and microglial BV2 cells, which all expressed mRNA for particular zinc transporters.
Conclusions/Significance
Taken together, comprehensive analysis is absolutely required for the demonstration of a variety of physiological and pathological responses mediated by Ca2+ in diverse cells enriched of Zn2+.

Link information
DOI
https://doi.org/10.1371/journal.pone.0127421
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/26010609
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000355183900089&DestApp=WOS_CPL
ID information
  • DOI : 10.1371/journal.pone.0127421
  • ISSN : 1932-6203
  • Pubmed ID : 26010609
  • Web of Science ID : WOS:000355183900089

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