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Analysis of networking of DNA repair in DNA repairdeficient mice with impaied spermatogenesis

精子发生受损的 DNA 修复缺陷小鼠 DNA 修复网络分析

基本信息

批准号：
18591782
负责人：
NAKANE Hironobu
金额：
$ 2.43万
依托单位：
Tottori University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591782/
关键词：
DNA reapor deficient mice genetical instability spermatogenesis networking of DNA repair

项目摘要

We focused on the relationship between networking of DNA repair and DNA repair-deficient mice with impaired spermatogenesis.DNA microarray analysis of testis in DNA repair deficient mice showed a different expression pattern of the base excision repair (BER), nucleotide excision repair (NER), translesion DNA synthesis (TLS), mismatch repair (MMR) compared to control. Now, we are confirming these different expression pattern by carrying out RT-PCR (reverse transcription PCR), immunostaining, western blotting, real-time PCR. In a preliminary study, we found the experimental condition that treated DNA repair deficient mice had abnormal spermatogenesis m their testes. So, we are trying to confirm these data, and carrying out further analysis.In addition, we found that XPG deficient mice (NER defect) had immature testes and ovaries, than were cheking time-specific gene expression in these organs. As we also found specific stain of platinum-blue in transmission electron microscopy, we were analyzing testes in DNA repair-deficient mice by transmission electron microscopy with platinum-blue stain. We can understand the relationship between networking of DNA repair and DNA repair-deficient mice with impaired spermatogenesis with these experiment.

我们重点关注DNA修复网络与精子发生受损的DNA修复缺陷小鼠之间的关系。DNA修复缺陷小鼠睾丸的DNA微阵列分析显示，与对照相比，碱基切除修复（BER）、核苷酸切除修复（NER）、跨损伤DNA合成（TLS）、错配修复（MMR）的表达模式不同。现在，我们正在通过进行RT-PCR（逆转录PCR）、免疫染色、蛋白质印迹、实时PCR来确认这些不同的表达模式。在一项初步研究中，我们发现经过治疗的 DNA 修复缺陷小鼠的实验条件下，其睾丸中的精子发生异常。因此，我们正在试图证实这些数据，并进行进一步的分析。此外，我们通过检查这些器官中的时间特异性基因表达，发现XPG缺陷小鼠（NER缺陷）的睾丸和卵巢不成熟。由于我们还在透射电子显微镜中发现了铂蓝的特异性染色，因此我们通过铂蓝染色的透射电子显微镜分析了 DNA 修复缺陷小鼠的睾丸。通过这些实验，我们可以了解DNA修复网络与精子发生受损的DNA修复缺陷小鼠之间的关系。