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Protein synthesis patterns in nuclear transfer, parthenogenetically activated and IVF bovine embryos during early development
Theriogenology
361
PROTEIN SYNTHESIS PATTERNS IN NUCLEAR TRANSFER, PARTHENOGENETICALLY ACl-IVATED AND IVF BOVINE EMBRYOS DURING EARLY DEVELOPMENT Z. Yang, A. K. Goff and L. C. Smith CRRA, Faculd de medecine veterinaire, Universite de Montreal, St-Hyacinthe, PQ, Canada J2S 7C6 Nuclear transfer has become an important tool in research both for fundamental and practical aspects of mammalian embryo development. Nuclei derived from transcriptionally activated embryos are able, in some cases, to support full term development after transfer to an enucleated oocyte suggesting complete reprogramming. However, little is known about the mechanism by which this occurs. The objective of this study was to investigate the reprogramming by analyzing quantitative and qualitative changes in protein synthesis in nuclear transfer (NT) embryos and to compare the changes observed in IVF and parthenogenetically activated (PA) embryos during early development. Bovine oocytes were matured for 24 h in v&-o, selected for the presence of a polar body, and randomly divided into 3 groups. NT oocytes were enucleated at 24 to 28 h after the onset of maturation, incubated for an additional 20 h and fused with a single blastomere from a 32-cell embrvo at 48 h after the onset of maturation. Oocvtes in the PA group were activated using an electric pulse with the same settings as the NT group. Oocvtes in the IVF group were fertilized in Fert-TALP (Parrish et al., 1988. Biol. Reurod. 38:1171-1180). All embryos from l- to 32-cell stages in the’3 groups were incubated with $%Imethionine (Amershan) for 2 h. Individual embryos were then assessed for uptake and incorporation, as well as protein synthesis patterns. Analysis of variance for uptake and incorooration was conducted using the comouter urocram of JMP (Version 2. SAS Institue Inc.. Gary: NC). Autoradiograms from-protein gels were >nalyzed using a laser densitometer (Ultra Scan XL, Pharmacier, Sweden) and the comouter program (GelScanXL Software Package. Pharmacia). The rate of uptake and incorporation of m
361
PROTEIN SYNTHESIS PATTERNS IN NUCLEAR TRANSFER, PARTHENOGENETICALLY ACl-IVATED AND IVF BOVINE EMBRYOS DURING EARLY DEVELOPMENT Z. Yang, A. K. Goff and L. C. Smith CRRA, Faculd de medecine veterinaire, Universite de Montreal, St-Hyacinthe, PQ, Canada J2S 7C6 Nuclear transfer has become an important tool in research both for fundamental and practical aspects of mammalian embryo development. Nuclei derived from transcriptionally activated embryos are able, in some cases, to support full term development after transfer to an enucleated oocyte suggesting complete reprogramming. However, little is known about the mechanism by which this occurs. The objective of this study was to investigate the reprogramming by analyzing quantitative and qualitative changes in protein synthesis in nuclear transfer (NT) embryos and to compare the changes observed in IVF and parthenogenetically activated (PA) embryos during early development. Bovine oocytes were matured for 24 h in v&-o, selected for the presence of a polar body, and randomly divided into 3 groups. NT oocytes were enucleated at 24 to 28 h after the onset of maturation, incubated for an additional 20 h and fused with a single blastomere from a 32-cell embrvo at 48 h after the onset of maturation. Oocvtes in the PA group were activated using an electric pulse with the same settings as the NT group. Oocvtes in the IVF group were fertilized in Fert-TALP (Parrish et al., 1988. Biol. Reurod. 38:1171-1180). All embryos from l- to 32-cell stages in the’3 groups were incubated with $%Imethionine (Amershan) for 2 h. Individual embryos were then assessed for uptake and incorporation, as well as protein synthesis patterns. Analysis of variance for uptake and incorooration was conducted using the comouter urocram of JMP (Version 2. SAS Institue Inc.. Gary: NC). Autoradiograms from-protein gels were >nalyzed using a laser densitometer (Ultra Scan XL, Pharmacier, Sweden) and the comouter program (GelScanXL Software Package. Pharmacia). The rate of uptake and incorporation of m