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. 2014 Mar;31(3):313-21.
doi: 10.1007/s10815-013-0159-y. Epub 2013 Dec 21.

Intracellular glutathione content, developmental competence and expression of apoptosis-related genes associated with G6PDH-activity in goat oocyte

Amir Hossein Abazari-Kia  1 , Abdollah Mohammadi-Sangcheshmeh, Maryam Dehghani-Mohammadabadi, Fatemeh Jamshidi-Adegani, Arash Veshkini, Mahdi Zhandi, Mehmet Ulas Cinar, Mohammad Salehi
Affiliations

Intracellular glutathione content, developmental competence and expression of apoptosis-related genes associated with G6PDH-activity in goat oocyte

Amir Hossein Abazari-Kia et al. J Assist Reprod Genet. 2014 Mar.

Abstract

Purpose: To associate glucose-6-phosphate dehydrogenase (G6PDH) activity in goat oocytes with intracellular glutathione (GSH) content, meiotic competence, developmental potential, and relative abundance of Bax and Bcl-2 genes transcripts.

Methods: Goat oocytes were exposed to brilliant cresyl blue (BCB) staining test and categorized into BCB(+) (blue-cytoplasm), and BCB(-) (colorless-cytoplasm) groups. A group of oocytes were not exposed to BCB test and was considered as a control group. After maturation in vitro, a group of oocytes were used for determination of nuclear status and intracellular GSH content while another group was subjected to parthenogenetic activation followed by in vitro embryo culture.

Results: We found that BCB(+) oocytes not only yielded higher rate of maturation, but also showed an increased level of intracellular GSH content than BCB(-) and control oocytes. Furthermore, BCB(+) oocytes produced more blastocysts than BCB(-) and control oocytes. Our data revealed that the expression of anti-apoptotic (Bcl-2) and pro-apoptotic (Bax) genes were interacted with G6PDH-activity in mature oocyte, their surrounding cumulus cells, and blastocyst-stage embryos.

Conclusions: The results of this study demonstrate that selection of goat oocytes based on G6PDH-activity through the BCB test improves their developmental competence, increases intracellular GSH content, and affects the expression of the apoptosis-related genes.

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Figures

Fig. 1
Fig. 1
Goat oocytes stained with Hoechst 33258 and evaluated under epifluorescence microscope, demonstrating oocytes in GV (a), GVBD (b), MI (c), and MII (d) stages
Fig. 2
Fig. 2
Effect of glucose-6-phosphate dehydrogenase (G6PDH) activity in goat oocytes on interacellular GSH level after in vitro maturation (IVM). BCB brilliant cresyl blue. a,b Bars with different letters represent groups that are significantly different (P < 0.05)
Fig. 3
Fig. 3
Relative mRNA expression of Bax and Bcl-2 genes for mature oocytes selected on their glucose-6-phosphate dehydrogenase (G6PDH) activity. BCB brilliant cresyl blue. * Asterisk shows significant difference between the groups (P < 0.05)
Fig. 4
Fig. 4
Relative mRNA expression of Bax and Bcl-2 genes for cumulus cells of mature oocytes selected on their glucose-6-phosphate dehydrogenase (G6PDH) activity. BCB brilliant cresyl blue. * Asterisk shows significant difference between the groups (P < 0.05)
Fig. 5
Fig. 5
Relative mRNA expression of Bax and Bcl-2 genes for blastocysts derived from oocytes that were selected on their glucose-6-phosphate dehydrogenase (G6PDH) activity. BCB brilliant cresyl blue. * Asterisk shows significant difference between the groups (P < 0.05)
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