title: Effects of chemical activation treatment on development of swamp buffalo (Bubalus bubalis) oocytes matured in vitro and fertilized by intracytoplasmic sperm injection.
abstract: The objective of this study was to optimize the activation protocol for buffalo oocytes after intracytoplasmic sperm injection (ICSI). The release of the second polar body (PB) at 3, 6 and 9 h after ICSI of in-vitro matured oocytes activated either with 5 μm ionomycin (Io) or with 7% ethanol (EtOH) was preliminary examined. The highest rate of second PB extrusion occurred at 3 h of activation, and the second PB extrusion in EtOH group was significantly higher than that in Io group. Oocytes that extruded the second PB were selected and cultured either with 1.9 mm 6-dimethylaminopurine (6-DMAP) for 3 h or with 10 μg/ml cycloheximide (CHX) for 5 h. Significantly higher rate of oocytes formed 2 pronuclei in EtOH combined with CHX (EtOH + CHX) (62%) group compared to those of Io + CHX (42%) and EtOH + 6-DMAP (48%) groups (p<0.01) whereas Io + 6-DMAP group showed intermediate value (58%). Significantly higher blastocyst formation rates were obtained in Io + 6-DMAP (29%) and EtOH + CHX (24%) groups than in Io + CHX (6%) and EtOH + 6-DMAP (17%) groups. Our results indicate that buffalo ICSI oocytes are effectively activated by combination treatment of Io with 6-DMAP and EtOH with CHX resulting in the highest cleavage and blastocyst formation rates.
authors.affiliation: Embryo Technology and Stem Cell Research Center and School of Biotechnology, Suranaree University of Technology, Nakhon Ratchasima, Thailand.
authors.forename: Y Y
authors.initials: YY
authors.lastname: Liang
authors.forename: D N
authors.initials: DN
authors.lastname: Ye
authors.forename: C
authors.initials: C
authors.lastname: Laowtammathron
authors.forename: T
authors.initials: T
authors.lastname: Phermthai
authors.forename: T
authors.initials: T
authors.lastname: Nagai
authors.forename: T
authors.initials: T
authors.lastname: Somfai
authors.forename: R
authors.initials: R
authors.lastname: Parnpai
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002020:Buffaloes; D002452:Cell Count; D002478:Cells, Cultured; D002970:Cleavage Stage, Ovum / Q000502:physiology; D003513:Cycloheximide / Q000494:pharmacology; D046149:Embryo Culture Techniques / Q000662:veterinary; D000431:Ethanol / Q000494:pharmacology; D005260:Female; D015759:Ionomycin / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects* / Q000502:physiology* / Q000648:ultrastructure; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary*
chemical_list: D047428:Protein Kinase Inhibitors; D000431:Ethanol; D015759:Ionomycin; C001020:N(6),N(6)-dimethyladenine; D003513:Cycloheximide; D000225:Adenine