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Noninvasive chemical enucleation of mouse oocytes
Author(s) -
Fulka Josef,
Moor Robert M.
Publication year - 1993
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080340412
Subject(s) - biology , enucleation , oocyte , microbiology and biotechnology , evolutionary biology , embryo , genetics
Abstract 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. © 1993 Wiley‐Liss, Inc.