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Korean Journal of Fertility and Sterility 2003;30(4):269-280.
Published online December 1, 2003.
The effect of Ni2+ on the intracellular Ca2+ increase of the mouse early 2-cell embryos.
Sook Young Yoon, Eun Mi Lee, In Ha Bae
Department of Biology, College of Natural Sciences, Sungshin Women's University, Korea.
Abstract
OBJECTIVE
We reported the overcoming effect of Ni2+ on the in vitro 2-cell block of mouse embryos. In this study, we aim to investigate whether Ni2+ should induce intracellular Ca2+ transient in the mouse embryos. MATERIALS AND METHODS: Embryos were collected at post hCG 32hr from the oviduct of the ICR mouse and cultured in M2 medium omitted phenol red. Intracellular Ca2+ was checked by using a confocal laser scanning microscope and fluo-3AM by using various intracellular Ca2+ antagonists. RESULTS: In 1mM Ni2+ treated medium which contained Ca2+(1.71mM), 75.7% of the embryos showed [Ca2+]i transient about 200 sec later. In the Ca2+-free medium, 69.8% of the embryos showed [Ca2+]i transient. In U73122, phospholipaseC(PLC) inhibitor (5uM, 10min) pretreated group, 33.3% of the embryos showed [Ca2+]i transient. Heparine, inositol 1,4,5-triphosphate receptor(IP3R) antagonist preinjected embryos showed no response with 1mM Ni2+. In danthrolene treatment, ryanodine receptor(RyR)-antagonist, 43% embryos showed [Ca2+]i transient but they showed delayed response about 340sec in the presence of Ca2+. CONCLUSIONS: Summing up the above results, Ni2+ seems to induce Ca2+-release from the Ca2+-store even in the Ca2+-free medium. IP3 receptors of the mouse 2-cell embryos might have an essential role for the intracellular Ca2+ increase by Ni2+.
Key Words: mouse in vitro 2-cell block; intracellualr Ca2+ increase; Ni2+; IP3 antagonist; ryanodine receptor antagonist
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