Development of parthenogenetic rat embryos.
In an effort to establish cloning technology for the rat, we tested several methods (electric stimulation, treatment with ethanol or strontium) for the parthenogenetic activation of rat oocytes. We observed marked individual differences among rats of the outbred Wistar strain in their ability to yield activatable oocytes. These differences were independent of the activation protocol and may be due to a genetic predisposition that is crucial for the parthenogenetic activation of oocytes. The activation of oocytes was dependent upon the time between superovulation of the donor animal and the collection of the embryos. Aged oocytes (derived about 24 h after superovulation) were more prone to activation by each method than were younger oocytes, and some even underwent spontaneous activation without treatment and exhibited pronuclear formation and blastocyst development. All activation methods were effective in generating parthenogenetic rat embryos, and rat parthenotes developed until implantation. However, in general, short-term (15 min) and long-term (2 h) strontium treatment was superior to stimulation by ethanol or electric pulse for parthenogenetic activation. These results will be helpful in achieving successful cloning in the rat.
Krivokharchenko
Alexander
A
Max-Delbrück Center for Molecular Medicine, D-13092 Berlin-Buch, Germany.
Popova
Elena
E
Zaitseva
Ioulia
I
Vil'ianovich
Larissa
L
Ganten
Detlev
D
Bader
Michael
M
D000818:Animals; D019976:Cloning, Organism / Q000379:methods*; D004558:Electric Stimulation; D000431:Ethanol / Q000494:pharmacology; D005260:Female; D008297:Male; D018613:Microscopy, Confocal / Q000662:veterinary; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D010312:Parthenogenesis / Q000502:physiology*; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar / Q000196:embryology / Q000502:physiology*; D013324:Strontium / Q000494:pharmacology*
D000431:Ethanol; D013324:Strontium