Our objective is to understand the function and mechanism of action of the first major secreted protein of the ovine conceptus, ovine trophoblast protein-1 (oTP-1) and the immunologically related bovine protein bTP-1. These proteins have been implicated in the process of maternal recognition of pregnancy in the ewe and cow. oTP-1 is produced between D 13 and 21 of pregnancy and maximally in the D 15-17 period. It appears to interact with the epithelial cells of the maternal uterus. We intend to improve the efficiency of oTP-1 purification by high performance chromatographic techniques and purify the immunologically cross-reacting bovine TP-1 for the first time. The relationships between the isoforms of the two proteins will be investigated by peptide mapping and by using monoclonal antibodies. We shall determine whether total conceptus secretory proteins, from which oTP-1 has been removed by immunoadsorption, can extend the lifespan of the corpus luteum when they are infused into the uterus of nonpregnant recipients. We shall also study whether the endometrial proteins which are induced by oTP-1 are produced during pregnancy. In the second part of the project, the main objective is to use recombinant DNA techniques for studying the structure, biosynthesis and function of oTP-1 and the related bTP-1 of the cow. We shall attempt to isolate """"""""full length"""""""" cDNA's to oTP-1 and bTP-1 from cDNA Libraries in the plasmid vector pUC-18 (already established). The nucleotide sequences of these cDNA's will be determined and primary amino acid sequences inferred. These sequences will be compared for homology with other known proteins and polypeptide hormones. The kinds of mRNA and their quantities will be evaluated in conceptuses during the period of maternal recognition of pregnancy. A final objective is to produce oTP-1 in quantity in a bacterial or mammalian cell system with the view to obviating the need for culturing large numbers of conceptuses. Longer term goals will be to determine whether conceptuses of other species produce oTP-1-like proteins and to introduce the full length genes for oTP-1 into fertilized eggs, with the view to producing transgenic animals which may have increased fertility.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
1R01HD021896-01
Application #
3321034
Study Section
Reproductive Biology Study Section (REB)
Project Start
1986-07-01
Project End
1990-06-30
Budget Start
1986-07-01
Budget End
1987-06-30
Support Year
1
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Missouri-Columbia
Department
Type
Earth Sciences/Resources
DUNS #
112205955
City
Columbia
State
MO
Country
United States
Zip Code
65211
Ezashi, Toshihiko; Imakawa, Kazuhiko (2017) Transcriptional control of IFNT expression. Reproduction 154:F21-F31
Green, Mark P; Harvey, Alexandra J; Spate, Lee D et al. (2016) The effects of 2,4-dinitrophenol and d-glucose concentration on the development, sex ratio, and interferon-tau (IFNT) production of bovine blastocysts. Mol Reprod Dev 83:50-60
Gupta, Rangan; Ezashi, Toshihiko; Roberts, R Michael (2012) Squelching of ETS2 transactivation by POU5F1 silences the human chorionic gonadotropin CGA subunit gene in human choriocarcinoma and embryonic stem cells. Mol Endocrinol 26:859-72
Bermejo-Alvarez, Pablo; Rosenfeld, Cheryl S; Roberts, R Michael (2012) Effect of maternal obesity on estrous cyclicity, embryo development and blastocyst gene expression in a mouse model. Hum Reprod 27:3513-22
Bermejo-Alvarez, P; Roberts, R M; Rosenfeld, C S (2012) Effect of glucose concentration during in vitro culture of mouse embryos on development to blastocyst, success of embryo transfer, and litter sex ratio. Mol Reprod Dev 79:329-36
Ezashi, Toshihiko; Matsuyama, Haruyo; Telugu, Bhanu Prakash V L et al. (2011) Generation of colonies of induced trophoblast cells during standard reprogramming of porcine fibroblasts to induced pluripotent stem cells. Biol Reprod 85:779-87
Zhou, Liang; Wang, Wei; Liu, Yongqing et al. (2011) Differentiation of induced pluripotent stem cells of swine into rod photoreceptors and their integration into the retina. Stem Cells 29:972-80
Schulz, Laura C; Roberts, R Michael (2011) Dynamic changes in leptin distribution in the progression from ovum to blastocyst of the pre-implantation mouse embryo. Reproduction 141:767-77
Roberts, R Michael; Fisher, Susan J (2011) Trophoblast stem cells. Biol Reprod 84:412-21
Roberts, R Michael; Katayama, Mika; Magnuson, Scott R et al. (2011) Transcript profiling of individual twin blastomeres derived by splitting two-cell stage murine embryos. Biol Reprod 84:487-94

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