Nucleus transfer in mammals: noninvasive approaches for the preparation of cytoplasts.
The production of cloned animals is a difficult and complex procedure that requires two basic steps. First, the cytoplast must be prepared by the enucleation of metaphase II oocytes. Second, the nucleus is transferred either by fusion or by direct microinjection into the cytoplast. The preparation of cytoplasts is a crucial step because they must be able to reprogram the transferred nucleus and to secure the development of reconstructed embryos. Moreover, the classical mechanical enucleation of metaphase II oocytes is rather technically difficult, requiring good equipment and considerable micromanipulation skill. For this reason the simplification of this step is permanently in the centre of interest of those scientists who are involved in the production of clones.
Fulka
Josef
J
Institute of Animal Production, POB 1, Prague 10, Czech Republic. fulka.josef@vuzv.cz
Loi
Pasqualino
P
Fulka
Helena
H
Ptak
Grazyna
G
Nagai
Taku
T
D000818:Animals; D000829:Animals, Domestic / Q000196:embryology / Q000235:genetics; D002499:Centrifugation, Density Gradient; D019976:Cloning, Organism / Q000379:methods*; D003513:Cycloheximide / Q000494:pharmacology; D003584:Cytological Techniques / Q000379:methods; D003703:Demecolcine / Q000494:pharmacology; D005047:Etoposide / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D008322:Mammals / Q000196:embryology / Q000235:genetics*; D016200:Maturation-Promoting Factor / Q000378:metabolism; D008677:Metaphase / Q000187:drug effects; D051379:Mice; D008846:Micromanipulation / Q000379:methods; D008856:Microscopy, Fluorescence; D008859:Microscopy, Polarization; D015739:Nocodazole / Q000494:pharmacology; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects; D008941:Spindle Apparatus / Q000187:drug effects / Q000378:metabolism; D013395:Sucrose / Q000494:pharmacology
D015759:Ionomycin; D013395:Sucrose; D005047:Etoposide; D003513:Cycloheximide; D016200:Maturation-Promoting Factor; D015739:Nocodazole; D003703:Demecolcine