玻璃化保存的小鼠窦前卵泡中DNA修复基因表达的评估
Assessment of DNA Repair Gene Expressions in Vitrified Mouse Preantral Follicles.
作者信息
Khodavandpour Zahra, Zavareh Saeed, Farrokh Parisa, Nasiri Meysam
机构信息
Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran.
School of Biology, Damghan University, Damghan, Iran. Electronic Address:
出版信息
Cell J. 2020 Jul;22(Suppl 1):81-88. doi: 10.22074/cellj.2020.6865. Epub 2020 Jul 18.
OBJECTIVE
Vitrification of the ovarian tissue is one of the techniques recommended for preserving the fertility of women who are dealing with infertility. Despite its benefits, our information about the molecular aspects of ovarian follicles vitrification is somehow ambiguous. Therefore, the aim of this study was to evaluate the expression pattern of DNA repair genes in vitrified preantral follicles.
MATERIALS AND METHODS
In this experimental study, the isolated preantral follicles (n=906) from 14-16 days old mice (n=12) were divided into three groups: fresh, toxic and vitrified which were cultured for 12 days. Preantral follicles were vitrified using cryotop followed by exposure to equilibration solution for five minutes and vitrification solution (VS) for 30 seconds. In the toxic group, preantral follicles were only placed in equilibration and vitrification media and they were then placed in the warming solutions without exposure to liquid nitrogen. On the second and sixth days of the culture period, real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was carried out to evaluate expression of the selected genes involved in DNA repair, including (MutS homolog 6), (Meiotic recombination 11), (Breast cancer type 1), (RAD51 recombinase), Pcna (Proliferating cell nuclear antigen) and (ATM serine/threonine kinase). In addition, developmental parameters including growth, survival rate, antrum cavity formation and ovulation were analyzed.
RESULTS
The relative mRNA expression of and on the second and sixth days of the culture period in vitrified group was significantly higher than those of the control and toxic groups, but there was no significant difference between the toxic and control groups. In addition, developmental parameters of follicles were similar in both toxic and control groups, while both were significantly higher than that of vitrified group.
CONCLUSION
Vitrification changes the expression pattern of DNA repair genes of the mouse preantral follicles.
目的
卵巢组织玻璃化是为处理不孕症的女性保留生育能力所推荐的技术之一。尽管有其益处，但我们对卵巢卵泡玻璃化的分子方面的信息在某种程度上仍不明确。因此，本研究的目的是评估玻璃化前体卵泡中DNA修复基因的表达模式。
材料与方法
在本实验研究中，从14 - 16日龄小鼠（n = 12）分离出的前体卵泡（n = 906）分为三组：新鲜组、毒性组和玻璃化组，培养12天。前体卵泡使用冷冻环进行玻璃化，随后暴露于平衡溶液5分钟和玻璃化溶液（VS）30秒。在毒性组中，前体卵泡仅置于平衡和玻璃化培养基中，然后置于复温溶液中，不暴露于液氮。在培养期的第二天和第六天，进行实时定量逆转录 - 聚合酶链反应（qRT - PCR）以评估参与DNA修复的选定基因的表达，包括（MutS同源物6）、（减数分裂重组11）、（乳腺癌1型）、（RAD51重组酶）、Pcna（增殖细胞核抗原）和（ATM丝氨酸/苏氨酸激酶）。此外，分析包括生长、存活率、卵泡腔形成和排卵在内的发育参数。
结果
玻璃化组培养期第二天和第六天的和的相对mRNA表达显著高于对照组和毒性组，但毒性组和对照组之间无显著差异。此外，毒性组和对照组卵泡的发育参数相似，而两者均显著高于玻璃化组。
结论
玻璃化改变了小鼠前体卵泡DNA修复基因的表达模式。
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