Cell allocation and chromosomal complement of parthenogenetic and IVF bovine embryos.
Considerable concerns exist regarding the quality of parthenogenetically activated embryos in terms of sufficient numbers of cells comprising the inner cell mass (ICM) and trophectoderm (TE) and the ploidy. Therefore, these two parameters were used to assess the quality of embryos derived from parthenogenetic activation by using calcium ionophore A23187 (CaI) followed by either 6-dimethylaminopurine (6-DMAP, 3.5 hr or 6.5 hr) or cycloheximide (CHX) plus cytochalasin D (CD). The conventional in vitro (IVF) produced embryos served as a control. Double staining of the parthenogenetic blastocysts showed that the total cell number (TC) of embryos from the 6-DMAP 3.5 hr (87.0 +/- 5.3) and CHX+CD (79.0 +/- 6.1) groups was not different (P > 0.05), but was lower than that of control embryos (116.0 +/- 5.8, P < 0.001). The mean ratios of inner cell mass (ICM) and trophectoderm (TE) cells in the 6-DMAP 3.5 hr group (0.57 +/- 0.04) and the control IVF group (0.50 +/- 0. 02) did not differ significantly. Both were higher than those of the CHX+CD group (0.36 +/- 0.02; P < 0.05). Further analysis of chromosomal compositions of developing stage embryos at day four after IVF or parthenogenetic activation demonstrated that prolonged treatment with 6-DMAP for 6.5 hr resulted in a significantly lower percentage of diploid embryos and a significantly higher percentage of abnormal ploidy embryos compared to treatment with 6-DMAP for 3.5 hr or with CHX and IVF. In conclusion, parthenogenetic activation of bovine oocytes with CaI followed by 6-DMAP for 3.5 hr could produce better quality embryos in terms of total cell numbers, the number of cells allocated to the ICM, and the ploidy of embryos.
Van De Velde
A
A
Department of Animal Science, University of Connecticut, Storrs, Connecticut, USA.
Liu
L
L
Bols
P E
PE
Ysebaert
M T
MT
Yang
X
X
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002417:Cattle; D002452:Cell Count / Q000187:drug effects; D002875:Chromosomes / Q000235:genetics*; D003513:Cycloheximide / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000187:drug effects*; D005307:Fertilization in Vitro; D009865:Oocytes / Q000187:drug effects; D010312:Parthenogenesis / Q000235:genetics*
D015638:Cytochalasin D; D000001:Calcimycin; C001020:N(6),N(6)-dimethyladenine; D003513:Cycloheximide; D000225:Adenine