Chemical activation with a combination of ionomycin and dehydroleucodine for production of parthenogenetic, ICSI and cloned bovine embryos.
The aim of this study was to evaluate the potential of dehydroleucodine (DhL), a new drug isolated from a medicinal herb used in Argentina, for activation of bovine oocyte. Several DhL concentrations and exposure times after ionomycin (Io) treatment were tested. The optimal DhL treatment, found for parthenogenetic development, was employed to produce bovine embryos by intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT). The best parthenogenic embryo developments were observed with 5 μM Io for 4 min followed by 5 μM DhL concentration and after 3-h exposure time (52.3% cleavage; 17.4% morulae; 7.3% blastocyst; n = 109). This treatment generated no significant differences with standard Io plus 6-dimethylaminopurine (DMAP) treatment in preimplantation embryo development. In our conditions, the embryo development reached after ICSI and SCNT assisted by the DhL treatment did not differ in terms of cleavage and blastocyst development from activation with standard Io plus DMAP treatment (p > 0.05). In conclusion, DhL utilization to activate oocytes and induce development of parthenogenotes, ICSI-embryos or SCNT-embryos is reported here for first time.
Vichera
G
G
Laboratorio de Biotecnología Animal, Facultad de Agronomía, Universidad de Buenos Aires, Buenos Aires, Argentina.
Alfonso
J
J
Duque
C C
CC
Silvestre
M A
MA
Pereyra-Bonnet
F
F
Fernández-Martín
R
R
Salamone
D
D
D000818:Animals; D002417:Cattle / Q000196:embryology*; D019976:Cloning, Organism; D004622:Embryo, Mammalian / Q000187:drug effects*; D047108:Embryonic Development / Q000187:drug effects; D015759:Ionomycin / Q000494:pharmacology*; D007783:Lactones / Q000737:chemistry / Q000494:pharmacology*; D015394:Molecular Structure; D053652:Nuclear Transfer Techniques / Q000662:veterinary; D010063:Ovum / Q000187:drug effects*; D010312:Parthenogenesis; D012717:Sesquiterpenes / Q000737:chemistry / Q000494:pharmacology*; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary
D007783:Lactones; D012717:Sesquiterpenes; C079949:dehydroleucodine; D015759:Ionomycin