Effects of Chemical Activation Treatment on Development of Swamp Buffalo (Bubalus bubalis) Oocytes Matured In Vitro and Fertilized by Intracytoplasmic Sperm Injection

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)...

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Bibliographic Details
Published in:Reproduction in domestic animals 2011-02, Vol.46 (1), p.e67-e73
Main Authors: Liang, YY, Ye, DN, Laowtammathron, C, Phermthai, T, Nagai, T, Somfai, T, Parnpai, R
Format: Article
Language:English
Subjects:
Adenine - analogs & derivatives
Adenine - pharmacology
Animal reproduction
Animals
Blastocyst - cytology
Blastocyst - physiology
Buffalo
Buffaloes
Cell Count
Cells, Cultured
Cellular biology
Cleavage Stage, Ovum - physiology
Cycloheximide - pharmacology
Embryo Culture Techniques - veterinary
Ethanol - pharmacology
Female
In vitro fertilization
Ionomycin - pharmacology
Oocytes - drug effects
Oocytes - physiology
Oocytes - ultrastructure
Protein Kinase Inhibitors - pharmacology
Sperm Injections, Intracytoplasmic - veterinary
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Summary: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.
ISSN:0936-6768
1439-0531
DOI:10.1111/j.1439-0531.2010.01636.x