| Background: Ammonium is produced in culture medium due to amino acids degradation
and has adverse effect on in vitro culture of embryo. In the current study, the
purpose was to evaluate the effects of ammuniom chloride (AC) on in vitro oocyte
maturation rate and early embryo development in the sheep and its effect on the expression
of Bcl-2.
Methods: In vitro maturation (IVM) was performed in the presence of various concentrations
(0, 29, 88,132,176 ƒÝM/ml) of ammonium chloride (NH4CL) (AC). Meiotic
maturation, embryonic development and expression of Bcl2 gene in Blastocyst
cells were determined. The data were analyzed by one-way ANOVA and Tukey post
HOC test, and values with p<0.05 were considered statistically significant.
Results: The highest concentration (176 £gM) of AC significantly decreased the rate
of fully expanded cumulus cells 24 hr after IVM compared with the control group
(p<0.05). Moreover, significantly lower rates of MII oocytes were found in the 176
£gM AC group compared with the 29 £gM AC group. The percentage of zygotes developing
to blastocysts in 176 £gM AC was lower than the other group. Also, supplementation
of the oocyte maturation media with 176 £gM AC decreased Bcl2 expression.
Conclusion: Our results suggested that significant increase in IVM rate could be obtained
with supplementation maturation medium with AC in a dose dependent manner.
Increased AC concentration led to lower blastocyst rate under normal condition.
However, regulation of pro¡Vapoptotic (Bcl-2) gene did not change with different
concentrations of AC supplementing. |