title: The effect of activation protocols on the development of cloned goat embryos.
abstract: 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.
authors.affiliation: The Bureau of Biotechnology for Animal Production, Department of Livestock Development, Pathumthani, 12000, Thailand.
authors.forename: Malee
authors.initials: M
authors.lastname: Apimeteetumrong
authors.forename: Anone
authors.initials: A
authors.lastname: Thuangsanthia
authors.forename: Narong
authors.initials: N
authors.lastname: Leingcharoen
authors.forename: Viboon
authors.initials: V
authors.lastname: Yiengvisavakul
authors.forename: Ayuth
authors.initials: A
authors.lastname: Harintharanon
authors.forename: Annop
authors.initials: A
authors.lastname: Kunavongkrit
authors.forename: Jureerat
authors.initials: J
authors.lastname: Sumretprasong
authors.forename: Xavier
authors.initials: X
authors.lastname: Vignon
authors.forename: Mongkol
authors.initials: M
authors.lastname: Techakumphu
mesh_terms: 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
chemical_list: D003571:Cytochalasin B; D000431:Ethanol; D015759:Ionomycin; C001020:N(6),N(6)-dimethyladenine; D000225:Adenine