The long-term objectives of this study are to define the roles of polypeptides secreted by the conceptus in embryo-maternal communication during the perimplantation period of pregnancy and to understand the underlying basis of the high rates of embryonic loss that occur around this time. The current proposal will focus largely on the involvement of embryo-derived interferons (IFN's) and other cytokines in triggering maternal responses to early sheep, cattle, pig and mouse embryos. The specific objectives are i) To determine the number of functional genes for the embryonic IFN'S, ovine trophoblast protein-1 (oTP-1) and the related bovine protein, bTP-1, by Southern blotting procedures with CDNA probes that will distinguish these IFN's from related IFN-alphas; ii) To map and sequence some of these genes and to define those regulatory regions that may be in common with other IFN-alphas Is and those that are uniquely associated with this embryonic class of IFN. A longer-term objective is to characterize transacting factors binding to these cis- control elements that are associated with oTP-1 and bTP-1 activation and repression in embryonic trophectoderm. iii) To produce large amounts of pure recombinant bTP-1 and oTP-1 in E. coli or cultured mammalian cells by recombinant DNA procedures. iv.) To test the physical, biochemical and biological properties of the recombinant proteins and, in particular, to assess their effects on a number of reproductive parameters, such as estrous cycle length, in sheep and cattle. v) To study how early bTP-1 or oTP-1 are inducible by double-stranded RNA in embryos and whether there are factors in uterine secretion that are responsible for the """"""""natural"""""""" induction of these IFN's around the time of maternal recognition of pregnancy. Induction will be studied by in situ hybridization procedures, and the relatively large numbers of bovine embryos necessary for these studies will be prepared by in vitro procedures from slaughterhouse-derived ovaries. vi) To characterize the secretory proteins responsible for IFN-like activity in pig and mouse embryos. It is hypothesized that production of IFN's by early embryos is a widespread phenomenon of broad significance to mammalian fertility. vii) A final long-term objective is to screen existing ovine libraries for cDNA's representing other proteins secreted by embryos around Day 15 of development, to sequence these cDNA's and to infer the likely functions of the proteins they represent. It is anticipated that the proteins comprise additional bioactive components involved in conceptus-maternal signalling during early pregnancy.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37HD021896-13
Application #
2673533
Study Section
Special Emphasis Panel (NSS)
Program Officer
Yoshinaga, Koji
Project Start
1986-07-01
Project End
2000-06-30
Budget Start
1998-07-01
Budget End
1999-06-30
Support Year
13
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Missouri-Columbia
Department
Veterinary Sciences
Type
Schools of Earth Sciences/Natur
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
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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
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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
Telugu, Bhanu Prakash V L; Ezashi, Toshihiko; Sinha, Sunilima et al. (2011) Leukemia inhibitory factor (LIF)-dependent, pluripotent stem cells established from inner cell mass of porcine embryos. J Biol Chem 286:28948-53

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