Although culture systems exist that support mouse oocyte maturation, fertilization, and preimplantation in vitro, there is still a large gap in our knowledge regarding the molecular bases of these processes. Moreover, such well defined In vitro culture systems are still lacking for other mammalian species. Four unresolved areas exist that relate directly to the ability of in vitro culture systems to support the processes of oocyte maturation, fertilization, and preimplantation development. First, the molecular basis for 'hardening' of the zona pellucida, which occurs during oocyte maturation in vitro and is correlated with the development of a metaphase II-arrested egg of reduced fertilizabilty, is still poorly understood.
The first aim of this grant is to purify from serum/follicular fluid the component(s) that inhibits modification of the zona pellucida during mouse oocyte maturation, to ascertain if the zona pellucida modification occurs during oocyte maturation in other species, and to determine if this serum/follicular fluid component(s) inhibits the modification of the zona pellucida in other species. Second, the regulation of the onset of embryonic gene transcription is poorly understood in any mammalian species.
The second aim of this grant is to identify proteins whose synthesis is a marker for transcriptional activation, and to define the role of protein phosphorylation in this process in other mammalian species, as well as to determine the effect of different culture media on embryonic gene transcription. Third, the function of endogenously produced growth factors by preimplantation embryos in development is not well-defined.
The third aim of this grant is to analyze the effect of different culture media on the expression of different growth factors and their receptors during preimplantation development of several mammalian species, and to determine biochemical and physiological responses of blastocysts to these growth factors. Last, the role of apical, trophectoderm protein secretion in implantation is poorly understood, and there is a paucity of information concerning any potential function of proteins that are secreted into the blastocoel in the regulation of Inner cell mass proliferation and differentiation.
The fourth aim of this grant is to analyze the effect of different culture media on the types of proteins secreted by blastocysts into the medium (apical) and into the blastocoel in several mammalian species. The comparative approach proposed in this grant will afford insights into common underlying biochemical and molecular mechanisms that regulate early development.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01HD028514-04
Application #
2201161
Study Section
Special Emphasis Panel (SRC (12))
Project Start
1991-09-01
Project End
1996-08-30
Budget Start
1994-09-01
Budget End
1995-08-31
Support Year
4
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Mitra, J; Schultz, R M (1996) Regulation of the acquisition of meiotic competence in the mouse: changes in the subcellular localization of cdc2, cyclin B1, cdc25C and wee1, and in the concentration of these proteins and their transcripts. J Cell Sci 109 ( Pt 9):2407-15
Ho, Y; Wigglesworth, K; Eppig, J J et al. (1995) Preimplantation development of mouse embryos in KSOM: augmentation by amino acids and analysis of gene expression. Mol Reprod Dev 41:232-8
Ning, X; Ward, C R; Kopf, G S (1995) Activation of a Gi protein in digitonin/cholate-solubilized membrane preparations of mouse sperm by the zona pellucida, an egg-specific extracellular matrix. Mol Reprod Dev 40:355-63
Ho, Y; Doherty, A S; Schultz, R M (1994) Mouse preimplantation embryo development in vitro: effect of sodium concentration in culture media on RNA synthesis and accumulation and gene expression. Mol Reprod Dev 38:131-41
Moos, J; Kalab, P; Kopf, G S et al. (1994) Rapid, nonradioactive, and quantitative method to analyze zona pellucida modifications in single mouse eggs. Mol Reprod Dev 38:91-3
Doherty, A S; Temeles, G L; Schultz, R M (1994) Temporal pattern of IGF-I expression during mouse preimplantation embryogenesis. Mol Reprod Dev 37:21-6
Eppig, J J; Schultz, R M; O'Brien, M et al. (1994) Relationship between the developmental programs controlling nuclear and cytoplasmic maturation of mouse oocytes. Dev Biol 164:1-9
Kalab, P; Visconti, P; Leclerc, P et al. (1994) p95, the major phosphotyrosine-containing protein in mouse spermatozoa, is a hexokinase with unique properties. J Biol Chem 269:3810-7
Anbari, K; Schultz, R M (1993) Effect of sodium and betaine in culture media on development and relative rates of protein synthesis in preimplantation mouse embryos in vitro. Mol Reprod Dev 35:24-8
Kalab, P; Schultz, R M; Kopf, G S (1993) Modifications of the mouse zona pellucida during oocyte maturation: inhibitory effects of follicular fluid, fetuin, and alpha 2HS-glycoprotein. Biol Reprod 49:561-7

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