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Creation and analysis of animal model monitoring abnormal cell replication

监测异常细胞复制的动物模型的创建和分析

基本信息

批准号：
16380194
负责人：
KON Yasuhiro
金额：
$ 10.24万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

项目摘要

DNA mismatch repair is a well-known system managing and controlling genomic structure that is conserved widely from bacteria to humans, and its mechanism is an action to substitute the gap of the genomic sequence into an accurate structure by the polymerase possessing proofreading activity. We found previously a unique phenotype in MRL mouse testis, i.e. metaphase-specific apoptosis (Msa). In addition, as a result of a genetics analysis of the testis, we found the mutation of Exonuclease 1(Exo1) as a factor associated with Msa. The purpose of this research project is to create and analyze the abnormal model mouse for the cell replication monitoring system paying attention to MRL mouse strain.In this research, we firstly clarified several unique phenotypes in MRL mice, i.e. the existence of heat stress resistant spermatocytes, the appearance of oocytes in the testis and the development of rete ovarii-derived ovarian cysts. Secondarily, the congenic mice carrying telomere region of the chromosome 1 in MRL mice were created and analyzed to confirm the re-appearance of Msa and autoimmune disease. Next, transgenic mice were created by introduction of antisense Exo1 and truncated Exo1 to examine whether the abnormality in the testis was derived from its volume or truncation. Finally, the exon skipping of Exo1 gene was examined in vitro, and the localization of Exo1 in the testis was clarified by immunohistochemistry using a specific antibody originally produced in the present study.These investigations suggest that many phenotypes presented in MRL mice are due to the abnormal activity monitoring cell replication, however, it is revealed that other factors independent on that might effect on some phenotypes in MRL mice. It would be further expected to use MRL mice as several unique models as well as an autoimmune disease.

DNA错配修复是一种广为人知的管理和控制基因组结构的系统，从细菌到人类都是如此，其机制是具有校对活性的聚合酶将基因组序列的缺口替换为准确结构的行为。我们以前在MRL小鼠睾丸中发现了一种独特的表型，即中期特异性凋亡(MSA)。此外，通过对睾丸的遗传学分析，我们发现外切酶1(Exo1)突变是MSA的一个相关因素。本研究旨在为关注MRL小鼠品系的细胞复制监测系统建立和分析异常模型小鼠。在本研究中，我们首先阐明了MRL小鼠特有的几种表型，即耐热应激精母细胞的存在、睾丸中卵母细胞的出现以及卵巢上皮来源的卵巢囊肿的发育。其次，建立携带MRL小鼠1号染色体端粒区的同源基因小鼠，并对其进行分析，以证实MSA和自身免疫性疾病的重现。接下来，通过引入反义Exo1和截短Exo1来建立转基因小鼠，以检查睾丸的异常是来自其体积还是来自于截断。最后，在体外检测了Exo1基因的外显子跳跃，并通过免疫组织化学方法明确了Exo1在睾丸中的定位。这些研究表明，MRL小鼠出现的许多表型是由于监测细胞复制的异常活动所致，但也揭示了其他独立于此的因素可能影响MRL小鼠的某些表型。人们还希望将MRL小鼠作为几种独特的模型和一种自身免疫性疾病。