title: Ultrastructural and karyotypic examination of in vitro produced bovine embryos developed in the sheep uterus.
abstract: This study examined whether development of bovine in vitro produced (IVP) blastocysts in the sheep uterus resulted in morphologically and karyotypically normal elongation stage bovine blastocysts. Seven day IVP bovine blastocysts, resulting from either in vitro maturation and fertilization, nuclear transfer (NT), or parthenogenic activation, were surgically transferred at the blastocyst stage into sheep uteri. Sheep were sacrificed after 7-9 days, and blastocysts were flushed from their uteri. One of each kind of IVP bovine blastocyst was recovered from sheep uteri for analysis by transmission electron microscopy, and nine NT blastocysts were used to establish cell cultures that were analysed for chromosome complement. TEM analysis of in vivo-derived elongation stage bovine and ovine blastocysts was done for comparative purposes. Most ultrastructural features of the 13-19 day blastocysts were similar to earlier stage blastocysts except that distinct alternative mitochondrial morphologies were found between epiblast and trophectoderm cells. Monociliated cells, presumably nodal cells, were observed in the bovine epiblast and hypoblast, and retrovirus-like particles were elaborated by cells in these same areas. Development in the sheep uterus of IVP bovine blastocysts resulted in the presence of crystalloid bodies in the trophectoderm cells, and apoptotic and necrotic cells were observed in the epiblast tissue. Thus, in vivo incubation in the sheep uterus allowed nearly normal development to the elongated blastocyst stage and may be useful for assessment of NT bovine blastocyst developmental competence. Cell cultures derived from the NT blastocysts had normal chromosome complements suggesting that activation by ionomycin and 6-dimethyl-aminopurine did not cause detrimental changes in ploidy in those blastocysts that developed.
authors.affiliation: USDA, ARS, LPSI, Gene Evaluation and Mapping Laboratory, Beltsville Agricultural Research Center, MD 20705, USA. ntalbot@lpsi.barc.usda.gov
authors.forename: N C
authors.initials: NC
authors.lastname: Talbot
authors.forename: A
authors.initials: A
authors.lastname: Powell
authors.forename: W
authors.initials: W
authors.lastname: Garrett
authors.forename: J L
authors.initials: JL
authors.lastname: Edwards
authors.forename: C
authors.initials: C
authors.lastname: Rexroad
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology / Q000648:ultrastructure; D002417:Cattle; D002999:Clone Cells; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000502:physiology* / Q000648:ultrastructure; D005314:Embryonic and Fetal Development; D005260:Female; D005307:Fertilization in Vitro; D007621:Karyotyping; D008297:Male; D008854:Microscopy, Electron; D053652:Nuclear Transfer Techniques; D010312:Parthenogenesis; D012756:Sheep / Q000502:physiology*