Diploid porcine parthenotes produced by inhibition of first polar body extrusion during in vitro maturation of follicular oocytes.
We investigated nuclear progression and in vitro embryonic development after parthenogenetic activation of porcine oocytes exposed to cytochalasin B (CB) during in vitro maturation (IVM). Nuclear progression was similar in control oocytes and oocytes matured in the presence of 1 microg/ml CB (IVM-CB group) by 37 h IVM; at this time the proportion of oocytes that had reached or passed through the anaphase-I stage did not differ significantly between the IVM-CB and the control groups (61.3 and 69.9% respectively; P < 0.05). After IVM for 37 h, no polar body extrusion was observed in the IVM-CB group. In these oocytes, the two lumps of homologous chromosomes remained in the ooplasm after their segregation and turned into two irregular sets of condensed chromosomes. By 41 h IVM, the double sets of chromosomes had reunited in 89.5% IVM-CB oocytes and formed a single large metaphase plate, whereas 68.8% of the control oocytes had reached the metaphase-II stage by this time. When IVM-CB oocytes cultured for 46 h were stimulated with an electrical pulse and subsequently cultured for 8 h without CB, 39.0% of them extruded a polar body and 82.9% of them had a female pronucleus. Chromosome analysis revealed that the majority of oocytes that extruded a polar body were diploid in both the control and the IVM-CB groups. However, the incidence of polyploidy in the IVM-CB group was higher than that in the control group (P < 0.05). In vitro development of diploid parthenotes in the control and the IVM-CB groups was similar in terms of blastocyst formation rates (45.8 and 42.8% respectively), number of blastomeres (39.9 and 44.4 respectively), the percentage of dead cells (4.3 and 2.9% respectively), and the frequency of apoptotic cells (7.3 and 6.3% respectively). Tetraploid embryos had a lower blastocyst formation rate (25.5%) and number of cells (26.2); however, the proportion of apoptotic nuclei (7.0%) was similar to that in diploid parthenotes. These results suggest that the proportion of homozygous and heterozygous genes does not affect in vitro embryo development to the blastocyst stage.
Somfai
Tamás
T
Reproductive Biology Unit, Department of Animal Science, National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan.
Ozawa
Manabu
M
Noguchi
Junko
J
Kaneko
Hiroyuki
H
Ohnuma
Katsuhiko
K
Karja
Ni Wayan Kurniani
NW
Fahrudin
Mokhamad
M
Maedomari
Naoki
N
Dinnyés
András
A
Nagai
Takashi
T
Kikuchi
Kazuhiro
K
D000818:Animals; D017209:Apoptosis; D001755:Blastocyst / Q000166:cytology; D002467:Cell Nucleus / Q000648:ultrastructure; D002478:Cells, Cultured; D002875:Chromosomes / Q000648:ultrastructure; D002970:Cleavage Stage, Ovum / Q000187:drug effects* / Q000648:ultrastructure; D003571:Cytochalasin B / Q000494:pharmacology*; D004171:Diploidy; D004558:Electric Stimulation; D047108:Embryonic Development; D005260:Female; D020287:In Situ Nick-End Labeling; D008297:Male; D009866:Oogenesis; D010312:Parthenogenesis; D013552:Swine
D003571:Cytochalasin B