Zn(2+), derived from cell preparation, partly attenuates Ca(2+)-dependent cell death induced by A23187, calcium ionophore, in rat thymocytes.
A23187, a calcium ionophore, is used to induce Ca(2+)-dependent cell death by increasing intracellular Ca(2+) concentration ([Ca(2+)](i)) under in vitro condition. Since this ionophore also increases membrane permeability of metal divalent cations such as Zn(2+) and Fe(2+) rather than Ca(2+), trace metal cations in cell suspension may affect Ca(2+)-dependent cell death induced by A23187. Therefore, the effects of chelators for divalent metal cations, EDTA and TPEN, on the A23187-induced cytotoxicity were cytometrically examined in rat thymocytes. The cytotoxicity of A23187 was attenuated by 1mM EDTA while it was augmented by 50 microM EDTA and 10 microM TPEN. These changes were statistically significant. The A23187-induced increase in Fluo-3 fluorescence intensity, a parameter for [Ca(2+)](i), was significantly reduced by 1mM EDTA while it was not the case for 50 microM EDTA and 10 microM TPEN. The intensity of FluoZin-3 fluorescence, a parameter for [Zn(2+)](i), increased by A23187 was respectively reduced by 50 microM EDTA and 10 microM TPEN. It is suggested that the attenuation of A23187-induced cytotoxicity by 1mM EDTA is due to the chelation of extracellular Ca(2+) and Zn(2+) while the augmentation by 50 microM ETDA or 10 microM TPEN is due to the chelation of extracellular Zn(2+). The Tyrode's solution without thymocytes contained 32.4 nM of zinc while it was 216.9 nM in the cell suspension. In conclusion, trace Zn(2+), derived from cell preparation, partly attenuates the Ca(2+)-dependent cell death induced by A23187.
Sakanashi
Yoko
Y
Laboratories of Cellular Signaling and Chemistry, Faculty of Integrated Arts and Sciences, The University of Tokushima, 1-1 Minami-zyosanjima, Tokushima 770-8502, Japan.
Oyama
Tomohiro M
TM
Matsuo
Yuki
Y
Oyama
Toshihisa B
TB
Nishimura
Yumiko
Y
Ishida
Shiro
S
Imai
Shoji
S
Okano
Yoshiro
Y
Oyama
Yasuo
Y
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D000001:Calcimycin / Q000494:pharmacology*; D017610:Calcium Compounds / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D004492:Edetic Acid / Q000494:pharmacology; D005029:Ethylenediamines / Q000494:pharmacology; D007476:Ionophores / Q000494:pharmacology*; D051381:Rats; D013950:Thymus Gland / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D017967:Zinc Compounds / Q000378:metabolism*
D017610:Calcium Compounds; D004338:Drug Combinations; D005029:Ethylenediamines; D007476:Ionophores; D017967:Zinc Compounds; D000001:Calcimycin; D004492:Edetic Acid; C044387:N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine