The effect of activation protocols on the development of cloned goat embryos.
This study was conducted to compare the developmental competence of somatic nuclear transfer (NT) embryos, after either ionomycin or ethanol activation, in locally bred goats. Donor cells were prepared from the ear skin fibroblasts of a female goat. Cells, at passage 3-8, starved by culturing in 0.5% FCS for 4-8 d, were used for NT. Immature oocytes were obtained from FSH-stimulated goats and matured for 22 hr before enucleation and NT. After fusion, the reconstructed embryos were activated with either ionomycin or ethanol followed by culturing in 6-dimethylaminopurine (6-DMAP) and cytochalasin B (CB), for 3 hr. In experiment I, the fused NT embryos (n=63, ionomycin and n=68, ethanol treatments, respectively) were cultured in B2 with a Vero co-culture system and their developmental competence was evaluated through to Day 9. In experiment II, the NT embryos at the 2-4 cell stage on Day 2 derived from each treatment (ionomycin n=46, and ethanol n=37), were transferred into 10 synchronous recipients. There were no significant differences between the NT embryos derived from the ionomycin and ethanol groups, in fusion (86.3% versus 82.9%), cleavage (90.5% versus 82.4%) and for morula/blastocyst development rates (9.5% versus 5.9%). Sixty percent (3/5) of the recipients from ionomycin became pregnant by midterm (2.5 mts) while only 20% (1/5) from ethanol treatment was pregnant by Day 45. The results demonstrate that activation with either ionomycin or ethanol in combination with 6-DMAP-CB treatment does not affect the development of cloned goat embryos.
Apimeteetumrong
Malee
M
The Bureau of Biotechnology for Animal Production, Department of Livestock Development, Pathumthani, 12000, Thailand.
Thuangsanthia
Anone
A
Leingcharoen
Narong
N
Yiengvisavakul
Viboon
V
Harintharanon
Ayuth
A
Kunavongkrit
Annop
A
Sumretprasong
Jureerat
J
Vignon
Xavier
X
Techakumphu
Mongkol
M
D000225:Adenine / Q000031:analogs & derivatives*; D000818:Animals; D019976:Cloning, Organism; D003571:Cytochalasin B; D004622:Embryo, Mammalian / Q000187:drug effects / Q000196:embryology*; D004627:Embryonic Induction / Q000187:drug effects*; D000431:Ethanol / Q000494:pharmacology*; D006041:Goats / Q000196:embryology*; D015759:Ionomycin / Q000494:pharmacology*; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000187:drug effects*; D013268:Stimulation, Chemical
D003571:Cytochalasin B; D000431:Ethanol; D015759:Ionomycin; C001020:N(6),N(6)-dimethyladenine; D000225:Adenine