Development of rabbit parthenogenetic oocytes and nuclear-transferred oocytes receiving cultured cumulus cells

• Published in: Theriogenology Vol. 54; no. 9; pp. 1469 - 1476
• Main Authors: Yin, X.J., Tani, T., Kato, Y., Tsunoda, Y.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-12-2000
• Subjects: activation; Adenine - analogs & derivatives; Adenine - pharmacology; Animals; Blastocyst - physiology; cumulus cells; Electric Stimulation; Embryo Transfer - veterinary; Enzyme Inhibitors - pharmacology; Female; nuclear transfer; Nuclear Transfer Techniques; Oocytes - physiology; Oocytes - ultrastructure; Ovary - cytology; Parthenogenesis; Pseudopregnancy; Rabbits; Superovulation; nuclear transfer; activation; cumulus cells
• ISSN: 0093-691X; 1879-3231
• DOI: 10.1016/S0093-691X(00)00468-4

The present study determined a suitable parthenogenetic activation procedure for rabbit oocytes and examined the developmental potential of enucleated oocytes receiving cultured cumulus cells. Unfertilized oocytes recovered from superovulated rabbits were activated with one or two sets of electrical pulses, with or without subsequent administration of 6-dimethylaminopurine (6-DMAP). The proportion of oocytes treated with one or two sets of electrical pulses and 6-DMAP that cleaved (87% and 98%, respectively) and developed into blastocysts (77% and 85%, respectively) was significantly higher (P < 0.05) than those activated with electrical pulses alone (30% and 42% for cleavage, 7% and 17% for blastocysts).
Cumulus cells separated from ovulated oocytes obtained from mature rabbits were cultured for three to five passages and then induced to quiescence by serum starvation before nuclear transfer. The enucleated oocytes receiving cumulus cells were activated with electrical pulses followed by the addition of 6-DMAP, and cultured in vitro for 5 to 6 d or transferred to pseudopregnant recipient females 1 d after activation. Of 186 nuclear-transferred oocytes, 123 (66%) cleaved and 42 (23%) developed into blastocysts. After transfer of 174 nuclear-transferred oocytes to 8 recipient females, a total of 3 implantation sites were observed in 3 recipient females but no fetuses were obtained.