title: Somatic cell nuclear transfer alters peri-implantation trophoblast differentiation in bovine embryos.
abstract: Abnormal placental development limits success in ruminant pregnancies derived from somatic cell nuclear transfer (SCNT), due to reduction in placentome number and consequently, maternal/fetal exchange. In the primary stages of an epithelial-chorial association, the maternal/fetal interface is characterized by progressive endometrial invasion by specialized trophoblast binucleate/giant cells (TGC). We hypothesized that dysfunctional placentation in SCNT pregnancies results from aberration in expression of genes known to be necessary for trophoblast proliferation (Mash2), differentiation (Hand1), and function (IFN-tau and PAG-9). We, therefore, compared the expression of these factors in trophoblast from bovine embryos derived from artificial insemination (AI), in vitro fertilization (IVF), and SCNT prior to (day 17) and following (day 40 of gestation) implantation, as well as TGC densities and function. In preimplantation embryos, Mash2 mRNA was more abundant in SCNT embryos compared to AI, while Hand1 was highest in AI and IVF relative to SCNT embryos. IFN-tau mRNA abundance did not differ among groups. PAG-9 mRNA was undetectable in SCNT embryos, present in IVF embryos and highest in AI embryos. In postimplantation pregnancies, SCNT fetal cotyledons displayed higher Mash2 and Hand1 than AI and IVF tissues. Allelic expression of Mash2 was not different among the groups, which suggests that elevated mRNA expression was not due to altered imprinting status of Mash2. The day 40 SCNT cotyledons had the fewest number of TGC compared to IVF and AI controls. Thus, expression of genes critical to normal placental development is altered in SCNT bovine embryos, and this is expected to cause abnormal trophoblast differentiation and contribute to pregnancy loss.
authors.affiliation: Centre de recherche en reproduction animale, Faculté de médecine vétérinaire, Université de Montréal, St-Hyacinthe, Québec, Canada J2S 7C6.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Arnold
authors.forename: Vilceu
authors.initials: V
authors.lastname: Bordignon
authors.forename: Réjean
authors.initials: R
authors.lastname: Lefebvre
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Murphy
authors.forename: Lawrence C
authors.initials: LC
authors.lastname: Smith
mesh_terms: D000483:Alleles; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D002417:Cattle; D002452:Cell Count; D002454:Cell Differentiation; D019976:Cloning, Organism; D046149:Embryo Culture Techniques; D010064:Embryo Implantation; D047108:Embryonic Development; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental; D005865:Gestational Age; D007315:Insemination, Artificial; D007371:Interferon-gamma / Q000235:genetics; D053652:Nuclear Transfer Techniques; D010920:Placenta; D010929:Placentation; D011247:Pregnancy; D011257:Pregnancy Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014327:Trophoblasts / Q000166:cytology*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D007371:Interferon-gamma; D011257:Pregnancy Proteins; D012333:RNA, Messenger; C097341:helix-loop-helix protein, eHAND