Inhibiting 3-phosphoglycerate kinase by EDTA stimulates the development of the cleavage stage mouse embryo

✍ Scribed by Michelle Lane; David K. Gardner

• Publisher: John Wiley and Sons
• Year: 2001
• Tongue: English
• Weight: 120 KB
• Volume: 60
• Category: Article
• ISSN: 1040-452X
• DOI: 10.1002/mrd.1083

✦ Synopsis

Abstract

Addition of EDTA to the medium significantly enhances mouse embryo development in culture. Embryos cultured in the absence of EDTA exhibit abnormal increases in glycolytic activity that result in reduced development. Culture with EDTA was able to prevent this increase in glycolysis and, therefore, maintain developmental competence. EDTA was shown to inhibit the activity of the glycolytic enzyme, 3‐phosphoglycerate kinase. Additionally, the effect of EDTA on maintaining high rates of embryo development in culture could be mimicked by the addition of Cibacron blue, an inhibitor of 3‐phosphoglycerate kinase. The inhibition of 3‐phosphoglycerate kinase by EDTA could be overcome by the addition of exogenous magnesium, indicating that the effect of EDTA was to reduce the availability of this co‐factor to the glycolytic kinases. Embryos cultured with EDTA had significantly lower levels of intracellular magnesium compared to embryos cultured without EDTA. Therefore, the effect of EDTA appears to be as a chelator of divalent cations such as magnesium, that are required for normal activity of kinases such as 3‐phosphoglycerate kinase. Mol. Reprod. Dev. 60: 233–240, 2001. © 2001 Wiley‐Liss, Inc.