The purpose of the proposed studies is to examine the interrelationships among amino acid transport systems in mouse oocytes, ova and preimplantation conceptuses and intra-and extracellular amino acid concentrations. The first and fourth aims are to complete identification of the transport systems present at each stage of development and to determine the transport systems present at each stage of development and to determine the location of systems in different cell types and membrane in blastocysts. The transport systems in granulosa cells will also be characterized in aim 4, and this data and that for oocytes will aid in determining how the latter cells are nourished during oogenesis.
The second aim i s to determine if the transport systems are subject to trans-inhibition or trans-stimulation (exchange) by determining if uptake or exodus of amino acids via each system is inhibited or stimulated, respectively, by substrateds of the system on the opposite side of the plasma membrane. Based on these results, mixtures of animo acids will be formulated either to maintain the amino avid levels in conceptuses at the intracellular concentrations of these substances normally found in situ or to raise or lower the intracellular concentrations of one or more amino acids. It will be determined in the third aim whether differences in the intracellular concentrations of amino acids, as produced in aim 2, are associated with differences in the abilities of preimplantation conceptuses to develop normally in vitro. It will also be determined if amino acids can affect the """"""""optimum"""""""" osmolarity at which preimplantation mouse conceptuses develop in vitro, since amino acids can be osmoregualtors and the """"""""optimum"""""""" osmolarity of standard media is below the osmolarity of secretions of the reproductive tract. Comparison of data from aims 1-4 to similar data for other species should provide insight into how early conceptuses of different species undergo similar morphological changes in widely different environments. Oocytes and early conceptuses o different species appear to express some of the same novel amino acid transport system activities at different stages of development. Such changes during evolution in the chronology of phenotypic expression are terms heterochrony, and elucidation of the mechanism controlling expression of these transport systems could help determine a mechanism of heterochrony. These studies are also important because it should be possible to enhance or inhibit fertility relatively innocuously based on the novel amino acid transport systems in eggs and preimplantation conceptuses.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD021801-05
Application #
3320937
Study Section
Human Embryology and Development Subcommittee 1 (HED)
Project Start
1986-04-01
Project End
1994-03-31
Budget Start
1990-04-01
Budget End
1991-03-31
Support Year
5
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Midwestern University
Department
Type
Schools of Osteopathy
DUNS #
City
Downers Grove
State
IL
Country
United States
Zip Code
60515
Van Winkle, L J; Dickinson, H R (1995) Differences in amino acid content of preimplantation mouse embryos that develop in vitro versus in vivo: in vitro effects of five amino acids that are abundant in oviductal secretions. Biol Reprod 52:96-104
Van Winkle, L J; Kakuda, D K; MacLeod, C L (1995) Multiple components of transport are associated with murine cationic amino acid transporter (mCAT) expression in Xenopus oocytes. Biochim Biophys Acta 1233:213-6
Van Winkle, L J; Patel, M; Wasserlauf, H G et al. (1994) Osmotic regulation of taurine transport via system beta and novel processes in mouse preimplantation conceptuses. Biochim Biophys Acta 1191:244-55
Van Winkle, L J; Campione, A L; Mann, D F et al. (1993) The cation receptor subsite of the choline transporter in preimplantation mouse conceptuses resembles a cation receptor subsite of several amino acid transporters. Biochim Biophys Acta 1146:38-44
Van Winkle, L J (1993) Endogenous amino acid transport systems and expression of mammalian amino acid transport proteins in Xenopus oocytes. Biochim Biophys Acta 1154:157-72
Van Winkle, L J; Mann, D F; Wasserlauf, H G et al. (1992) Mediated Na(+)-independent transport of L-glutamate and L-cystine in 1- and 2-cell mouse conceptuses. Biochim Biophys Acta 1107:299-304
Van Winkle, L J; Campione, A L (1992) Novel bumetanide-sensitive K+ transport in preimplantation mouse conceptuses. Am J Physiol 263:C773-9
Van Winkle, L J; Mann, D F; Weimer, B D et al. (1991) Na(+)-dependent transport of anionic amino acids by preimplantation mouse blastocysts. Biochim Biophys Acta 1068:231-6
Van Winkle, L J; Campione, A L (1991) Ouabain-sensitive Rb+ uptake in mouse eggs and preimplantation conceptuses. Dev Biol 146:158-66
Van Winkle, L J; Campione, A L; Gorman, J M (1990) Inhibition of transport system b0,+ in blastocysts by inorganic and organic cations yields insight into the structure of its amino acid receptor site. Biochim Biophys Acta 1025:215-24

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