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Amino acid carryover in the subzonal space of mouse fertilized ova affects subsequent transport kinetics

Published online by Cambridge University Press:  24 April 2009

Nirmala Rudraraju  and
Jay M. Baltz
Nirmala Rudraraju
Affiliation:
Ottawa Health Research Institute, Ottawa, Ontario, Canada. Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada.
Jay M. Baltz*
Affiliation:
Moses and Rose Loeb Research Centre, Ottawa Health Research Institute, 725 Parkdale Ave., Ottawa, Ontario K1Y 4E9, Canada. Ottawa Health Research Institute, Ottawa, Ontario, Canada. Department of Obstetrics and Gynecology, Division of Reproductive Medicine, University of Ottawa, Ottawa, Ontario, Canada. Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada.
*
All correspondence to: Jay M. Baltz. Moses and Rose Loeb Research Centre, Ottawa Health Research Institute, 725 Parkdale Ave., Ottawa, Ontario K1Y 4E9, Canada. Tel: +1 613 798 5555 ext. 13714. e-mail: jbaltz@ohri.ca
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Summary

We have investigated whether culture in glycine-containing medium affects subsequent glycine transport by the specific transport system, GLYT1, which is the sole glycine transporter in fertilized mouse ova. When fertilized ova were maintained for 6 h in culture with a physiological level of glycine (1 mM), subsequent transport of radiolabelled glycine was decreased by 40% compared with fertilized ova that had been maintained in glycine-free medium. Kinetic measurements showed that the apparent glycine affinity was decreased after culture with glycine (Km increased from 0.20 to 0.41 mM), but maximal transport rate was unchanged (similar Vmax of 20 and 23 fmol/fertilized ovum/min). These findings could have reflected activation of GLYT1 by prolonged substrate starvation, similar to some other amino acid transport systems. However, our findings were instead consistent with the alteration in glycine transport being due to trapping of glycine within the zona pellucida resulting in competitive transport inhibition even after ova were removed from glycine-containing media. First, even very brief exposures to glycine resulted in decreased subsequent glycine transport rates, with a maximal effect apparent within ~6 min. Second, extensive washing (at least six) reversed the effect. Third, the effect was absent when zona-free fertilized ova were used. Thus, it appears that components of the external environment of preimplantation embryos may continue to affect transport kinetics for a period even after embryos are removed from environments that contain them.

Keywords

Amino acidFertilized ovumGlycineTransportZona pellucida
Type
Research Article
Information
Zygote , Volume 17 , Issue 4 , November 2009 , pp. 281 - 287
Copyright
Copyright © Cambridge University Press 2009

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