Noninvasive chemical enucleation of mouse oocytes.
A noninvasive method of enucleating mouse oocytes has been developed and evaluated. Strong chromosome to chromosome binding was induced by culturing early metaphase I oocytes in etoposide supplemented medium. Subsequent expulsion of the entire chromosome complex during polar body extrusion was facilitated by exposing the etoposide treated oocytes to a combination of cycloheximide and etoposide during anaphase and telophase. This simple two-step chemical enucleation procedure yields fully enucleated mouse oocytes in 96% of cases. Chemically enucleated oocytes do not contain maturation promoting factor (MPF) at the end of etoposide-cycloheximide enucleation. MPF levels are, however, restored during subsequent incubation in drug-free medium and, after 15 h of post-enucleation culture, the cytoplasts regain their full capacity for parthenogenetic activation and nuclear remodelling. We believe that this novel enucleation technique will greatly facilitate the research in nuclear transplantation.
Fulka
J
J
Department of Molecular Embryology, AFRC Institute of Animal Physiology and Genetics Research, Babraham, Cambridge, United Kingdom.
Moor
R M
RM
D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000648:ultrastructure*; D003513:Cycloheximide / Q000494:pharmacology; D005047:Etoposide / Q000494:pharmacology; D005260:Female; D066298:In Vitro Techniques; D016200:Maturation-Promoting Factor / Q000378:metabolism; D008540:Meiosis; D000090204:Mesothelin; D051379:Mice; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000187:drug effects / Q000648:ultrastructure*
C000715528:Msln protein, mouse; D005047:Etoposide; D003513:Cycloheximide; D016200:Maturation-Promoting Factor; D000090204:Mesothelin