Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 Jul-Sep;17(3):144-50.

The Effect of Ammonium Chloride Concentration in In Vitro Maturation Culture on Ovine Embryo Development

Ali Golchin  1 , Reza Asadpour  1 , Leila Roshangar  2 , Raziallah Jafari-Jozani  1
Affiliations

The Effect of Ammonium Chloride Concentration in In Vitro Maturation Culture on Ovine Embryo Development

Ali Golchin et al. J Reprod Infertil. 2016 Jul-Sep.

Abstract

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 μM/ml) of ammonium chloride (NH 4 CL) (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 μM) 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 μM AC group compared with the 29 μM AC group. The percentage of zygotes developing to blastocysts in 176 μM AC was lower than the other group. Also, supplementation of the oocyte maturation media with 176 μM 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-apoptotic (Bcl-2) gene did not change with different concentrations of AC supplementing.

Keywords: Ammonium chloride; Gene expression; Ovine embryo.

PubMed Disclaimer

Figures

Figure 1.
Figure 1.
Cumulus cell expansion decreases following IUVM in presence of ammonium chloride
Figure 2.
Figure 2.
Oocyte staining and determination of the stage of nuclear maturation
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Alexiou M, Leese HJ. Purine utilisation, de novo synthesis and degradation in mouse preimplantation embryos. Development. 1992; 114( 1): 185– 92. - PubMed
    1. Butler WR, Calaman JJ, Beam SW. Plasma and milk urea nitrogen in relation to pregnancy rate in lactating dairy cattle. J Anim Sci. 1996; 74( 4): 858– 65. - PubMed
    1. Cory S, Adams JM. Matters of life and death: programmed cell death at Cold Spring Harbor. Biochim Biophys Acta. 1998; 1377( 2): R25– 44. - PubMed
    1. Edwards LJ, Williams DA, Gardner DK. Intracellular pH of the mouse preimplantation embryo: amino acids act as buffers of intracellular pH. Hum Reprod. 1998; 13( 12): 3441– 8. - PubMed
    1. Ellis RE, Yuan JY, Horvitz HR. Mechanisms and functions of cell death. Annu Rev Cell Biol. 1991; 7: 663– 98. - PubMed

LinkOut - more resources