Overexpression of Mitochondrial Genes (Mitochondrial Transcription Factor A and Cytochrome c Oxidase Subunit 1) in Mouse Metaphase II Oocytes following Vitrification via Cryotop

Document Type: Original Article(s)

Authors

1 Department of Biology, College of Science, Fars Science and Research Branch, Islamic Azad University, Fars, Iran; and Department of Biology, College of Science, Shiraz Branch, Islamic Azad University, Shiraz, Iran

2 Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; and Department of Medical Genetics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

3 Department of Biology, College of Science, Shiraz Branch, Islamic Azad University, Shiraz, Iran;

4 Stem Cells Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

5 Department of Biology, Payam Noor University, Tehran, Iran

10.30476/ijms.2019.44960

Abstract

Background: Gamete cryopreservation is an inseparable part of assisted reproductive technology, and vitrification is an effective approach to the cryopreservation of oocytes. The aim of this study was to investigate vitrification effects on the expression levels of mitochondrial transcription factor A (Tfam) and mitochondrial-encoded cytochrome c oxidase subunit 1 (Cox1) in mouse metaphase II oocytes.Methods: Oocytes were selected by simple random sampling and distributed amongst five experimental groups (control [n=126], docetaxel [n=132], docetaxel+cryoprotectant agent [CPA] [n=134], docetaxel+vitrification [n=132], and vitrification [n=123]). After the warming process, the oocytes were fertilized and cultured into a 2-cell stage. Then, the effects of vitrification on the expression of the Tfam and Cox1 genes were determined via real-time reverse transcriptase polymerase chain reaction. Each group was compared with the control group. The data were analyzed with ANOVA using GraphPad and SPSS, version 21.Results: A significant decrease was observed in the fertilization rate of each group in comparison with the control group (P=0.001). The rate of 2-cell formation after in vitro fertilization was significantly lower in both vitrification groups (docetaxel+vitrification and vitrification) than in the non-vitrification groups (fresh control and docetaxel) and control group (P=0.001 and P=0.004). The expression level of Cox1 was significantly higher in the vitrification group than in the control group (P=0.01), while it was lower in the docetaxel group than that in the control group (P=0.04). The expression level of the Tfam gene was significantly high in the vitrification group (vitrification+docetaxel) and the non-vitrified group (docetaxel+CPA) in comparison with the control group (P=0.01).Conclusion: This study indicated that the vitrification of mouse MII oocytes increased the expression of the Tfam and Cox1 genes.

Keywords