Hostname: page-component-78c5997874-t5tsf Total loading time: 0 Render date: 2024-11-09T15:25:58.320Z Has data issue: false hasContentIssue false
  • English
  • Français

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: [email protected]
Get access Rights & Permissions [Opens in a new window]

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

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Biggers, J.D., Whittingham, D.G. & Donahue, R.P. (1967). The pattern of energy metabolism in the mouse oocyte and zygote. Proc. Natl. Acad. Sci. USA 58, 560–7.CrossRefGoogle ScholarPubMed
Carayannopoulos, M.O., Chi, M.M., Cui, Y., Pingsterhaus, J.M., McKnight, R.A., Mueckler, M., Devaskar, S.U. & Moley, K.H. (2000). GLUT8 is a glucose transporter responsible for insulin-stimulated glucose uptake in the blastocyst. Proc. Natl. Acad. Sci. USA 97, 7313–8.CrossRefGoogle ScholarPubMed
Dandekar, P. & Talbot, P. (1992). Perivitelline space of mammalian oocytes: extracellular matrix of unfertilized oocytes and formation of a cortical granule envelope following fertilization. Mol. Reprod. Dev. 31, 135–43.CrossRefGoogle ScholarPubMed
Dawson, K.M. & Baltz, J.M. (1997). Organic osmolytes and embryos: substrates of the Gly and beta transport systems protect mouse zygotes against the effects of raised osmolarity. Biol. Reprod. 56, 1550–8.CrossRefGoogle ScholarPubMed
Dawson, K.M., Collins, J.L. & Baltz, J.M. (1998). Osmolarity-dependent glycine accumulation indicates a role for glycine as an organic osmolyte in early preimplantation mouse embryos. Biol. Reprod. 59, 225–32.CrossRefGoogle ScholarPubMed
Franchi Gazzola, R., Sala, R., Bussolati, O., Visigalli, R., Dall'Asta, V., Ganapathy, V. & Gazzola, G.C. (2001). The adaptive regulation of amino acid transport system A is associated to changes in ATA2 expression. FEBS Letts. 490, 11–14.CrossRefGoogle Scholar
Gardner, D.K. & Lane, M. (1996). Alleviation of the ‘2-cell block’ and development to the blastocyst of CF1 mouse embryos: role of amino acids, EDTA and physical parameters. Hum. Reprod. 11, 2703–12.CrossRefGoogle Scholar
Guerin, J.F., Gallois, E., Croteau, S., Revol, N., Maurin, F., Guillaud, J. & Menezo, Y. (1995). Techniques de récolte et aminogrammes des liquides tubaire et folliculaire chez les femelles domestiques. Revue Med. Vet. 146, 805–14.Google Scholar
Gwatkin, R.B. (1967). Passage of mengovirus through the zona pellucida of the mouse morula. J. Reprod. Fertil. 13, 577–8.CrossRefGoogle ScholarPubMed
Harding, E.A., Day, M.L., Gibb, C.A., Johnson, M.H. & Cook, D.I. (1999). The activity of the H+-monocarboxylate cotransporter during pre-implantation development in the mouse. Pflugers Arch. 438, 397–404.CrossRefGoogle ScholarPubMed
Harris, S.E., Gopichandran, N., Picton, H.M., Leese, H.J. & Orsi, N.M. (2005). Nutrient concentrations in murine follicular fluid and the female reproductive tract. Theriogenology 64, 992–1006.CrossRefGoogle ScholarPubMed
Lawitts, J.A. & Biggers, J.D. (1993). Culture of preimplantation embryos. Methods Enzymol. 225, 153–64.CrossRefGoogle ScholarPubMed
Leese, H.J. & Barton, A.M. (1984). Pyruvate and glucose uptake by mouse ova and preimplantation embryos. J. Reprod. Fertil. 72, 9–13.CrossRefGoogle ScholarPubMed
Legge, M. (1995). Oocyte and zygote zona pellucida permeability to macromolecules. J. Exp. Zool. 271, 145–50.CrossRefGoogle ScholarPubMed
Pastor-Anglada, M., Felipe, A., Casado, F.J., Ferrer-Martinez, A. & Gomez-Angelats, M. (1996). Long-term osmotic regulation of amino acid transport systems in mammalian cells. Amino Acids 11, 135–51.CrossRefGoogle ScholarPubMed
Phillips, D.M. & Shalgi, R.M. (1980). Surface properties of the zona pellucida. J. Exp. Zool. 213, 1–8.CrossRefGoogle ScholarPubMed
Steeves, C.L. & Baltz, J.M. (2005). Regulation of intracellular glycine as an organic osmolyte in early preimplantation mouse embryos. J. Cell. Physiol. 204, 273–9.CrossRefGoogle ScholarPubMed
Steeves, C.L., Hammer, M.A., Walker, G.B., Rae, D., Stewart, N.A. & Baltz, J.M. (2003). The glycine neurotransmitter transporter GLYT1 is an organic osmolyte transporter regulating cell volume in cleavage-stage embryos. Proc. Natl. Acad. Sci. USA 100, 13982–7.CrossRefGoogle ScholarPubMed
Tanaka, K., Yamamoto, A. & Fujita, T. (2005). Functional expression and adaptive regulation of Na+-dependent neutral amino acid transporter SNAT2/ATA2 in normal human astrocytes under amino acid starved condition. Neurosci. Letts. 378, 70–5.CrossRefGoogle ScholarPubMed
Turner, K. & Horobin, R.W. (1997). Permeability of the mouse zona pellucida: a structure-staining-correlation model using coloured probes. J. Reprod. Fertil. 111, 259–65.CrossRefGoogle ScholarPubMed
Van Winkle, L.J. (2001). Amino Acid Transport Regulation and Early Embryo Development. Biol. Reprod. 64, 1–12.CrossRefGoogle ScholarPubMed
Van Winkle, L.J., Haghighat, N., Campione, A.L. & Gorman, J.M. (1988). Glycine transport in mouse eggs and preimplantation conceptuses. Biochim. Biophys. Acta 941, 241–56.CrossRefGoogle ScholarPubMed