Abstract
Radiolabeled compounds that are substrates for transmembrane transporters can be used to study transport and metabolism in mammalian oocytes and preimplantation embryos. Because even very small amounts of radioisotopes can be detected, these techniques are feasible to use with only a few oocytes or embryos, even down to the level of single oocytes or embryos. Here, we describe the methods for determining the transport and accumulation of radiolabeled compounds into oocytes and preimplantation embryos and the determination of the rate of saturable transport via specific transporters in the plasma membrane.
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References
Van Winkle LJ, Campione AL (1991) Ouabain-sensitive Rb+  uptake in mouse eggs and preimplantation conceptuses. Dev Biol 146:158–166
Van Winkle LJ, Campione AL, Gorman JM, Weimer BD (1990) Changes in the activities of amino acid transport systems b0,+ and L during development of preimplantation mouse conceptuses. Biochim Biophys Acta 1021: 77–84
Van Winkle LJ, Campione AL, Farrington BH (1990) Development of system B0,+ and a broad-scope Na(+)-dependent transporter of zwitterionic amino acids in preimplantation mouse conceptuses. Biochim Biophys Acta 1025:225–233
Van Winkle LJ, Haghighat N, Campione AL, Gorman JM (1988) Glycine transport in mouse eggs and preimplantation conceptuses. Biochim Biophys Acta 941:241–256
Van Winkle LJ, Campione AL, Gorman JM (1988) Na+-independent transport of basic and zwitterionic amino acids in mouse blastocysts by a shared system and by processes which distinguish between these substrates. J Biol Chem 263:3150–3163
Van Winkle LJ, Patel M, Wasserlauf HG, Dickinson HR, Campione AL (1994) Osmotic regulation of taurine transport via system beta and novel processes in mouse preimplantation conceptuses. Biochim Biophys Acta 1191:244–255
Colonna R, Mangia F (1983) Mechanisms of amino acid uptake in cumulus-enclosed mouse oocytes. Biol Reprod 28:797–803
Haghighat N, Van Winkle LJ (1990) Developmental change in follicular cell-enhanced amino acid uptake into mouse oocytes that depends on intact gap junctions and transport system Gly. J Exp Zool 253:71–82
Higgins BD, Kane MT (2003) Inositol transport in mouse oocytes and preimplantation embryos: effects of mouse strain, embryo stage, sodium and the hexose transport inhibitor, phloridzin. J Reprod Fertil 125:111–118
Pelland AM, Corbett HE, Baltz JM (2009) Amino acid transport mechanisms in mouse oocytes during growth and meiotic maturation. Biol Reprod 81:1041–1054
Tartia AP, Rudraraju N, Richards T, Hammer MA, Talbot P, Baltz JM (2009) Cell volume regulation is initiated in mouse oocytes after ovulation. Development 136:2247–2254
Steeves CL, Hammer MA, Walker GB, Rae D, Stewart NA, Baltz JM (2003) The glycine neurotransmitter transporter GLYT1 is an organic osmolyte transporter regulating cell volume in cleavage-stage embryos. Proc Natl Acad Sci USA 100:13982–13987
Dawson KM, Collins JL, Baltz JM (1998) Osmolarity-dependent glycine accumulation indicates a role for glycine as an organic osmolyte in early preimplantation mouse embryos. Biol Reprod 59:225–232
Anas MK, Lee MB, Zhou C, Hammer MA, Slow S, Karmouch J, Liu XJ, Broer S, Lever M, Baltz JM (2008) SIT1 is a betaine/proline transporter that is activated in mouse eggs after fertilization and functions until the 2-cell stage. Development 135:4123–4130
Anas MKI, Hammer MA, Lever M, Stanton JA, Baltz JM (2007) The organic osmolytes betaine and proline are transported by a shared system in early preimplantation mouse embryos. J Cell Physiol 210:266–277
Van Winkle LJ (1999) Transport kinetics. In: Van Winkle LJ (ed) Biomembrane transport. Academic, San Diego, pp 65–131
Hammer MA, Kolajova M, Leveille M, Claman P, Baltz JM (2000) Glycine transport by single human and mouse embryos. Hum Reprod 15:419–426
Van Winkle LJ (2001) Amino acid transport regulation and early embryo development. Biol Reprod 64:1–12
Lawitts JA, Biggers JD (1993) Culture of preimplantation embryos. Methods Enzymol 225:153–164
Acknowledgements
These techniques as carried out in JMB’s laboratory were developed and used in projects that were funded by the Canadian Institutes of Health Research (CIHR) including operating grants and the Program on Oocyte Health. H.E.C. and S.R. were supported by scholarships from the CIHR Institute of Human Development, Child and Youth Health Training Program in Reproduction, Early Development and the Impact on Health (REDIH). We thank Ms. Megan Kooistra for helping validate and edit these protocols.
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Baltz, J.M., Corbett, H.E., Richard, S. (2013). Measuring Transport and Accumulation of Radiolabeled Substrates in Oocytes and Embryos. In: Homer, H. (eds) Mammalian Oocyte Regulation. Methods in Molecular Biology, vol 957. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-191-2_11
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DOI: https://doi.org/10.1007/978-1-62703-191-2_11
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-62703-191-2
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