Nystuen Arne M, Sachs Andrew J, Yuan Yang, Heuermann Laura, Haider Neena B
The Department of Genetics, Cell Biology and Anatomy, University of Nebraska Medical Center, Omaha, NE, 68198-5805, USA.
Mamm Genome. 2008 Sep;19(9):623-33. doi: 10.1007/s00335-008-9138-5. Epub 2008 Sep 3.
The nmf193 mutant was generated by a large-scale ENU mutagenesis screen and originally described as having a dominantly inherited phenotype characterized by fundus abnormalities. We determined that nmf193 mice exhibit outer-segment defects and progressive retinal degeneration. Clinical examination revealed retinal spotting apparent at 6 weeks of age. Histologic analysis of homozygous mutant mice at 6 weeks indicated an absence of outer segments (OS) and a 50% reduction of photoreceptor cells which progressed to complete loss of photoreceptors by 10 months. Mice heterozygous for the nmf193 mutation had a less severe phenotype of shortened outer segments at 2 months with progressive loss of photoreceptor cells to 50% by 10 months. A positional cloning approach using a DNA pooling strategy was performed to identify the causative mutation in nmf193 mice. The nmf193 mutation was linked to chromosome 17 and fine mapped to an interval containing the peripherin/rds (Prph2) gene. Mutation analysis identified a single base change in Prph2 that causes aberrant splicing between exons 1 and 2. Interestingly, a comparative histologic analysis demonstrated that Prph2 ( nmf193/+ ) mutants have similar photoreceptor degeneration to that of Nr2e3 ( rd7/rd7 ). We show that Prph2 mRNA and protein levels are reduced in the Nr2e3 ( rd7/rd7 ) mutant compared to control littermates. Chromatin immunoprecipitation analysis shows that Prph2 is a direct target of NR2E3. In addition, the downregulation of Prph2 gene expression is similar in both the Nr2e3 ( rd7/rd7 ) and Prph2 ( nmf193/+ ) mutants, suggesting that the reduction of Prph2 may contribute to the degenerative pathology seen in Nr2e3 ( rd7/rd7 ).
nmf193突变体是通过大规模ENU诱变筛选产生的,最初被描述为具有以眼底异常为特征的显性遗传表型。我们确定nmf193小鼠表现出外段缺陷和进行性视网膜变性。临床检查显示在6周龄时出现明显的视网膜斑点。对6周龄纯合突变小鼠的组织学分析表明,外段(OS)缺失,光感受器细胞减少50%,到10个月时光感受器完全丧失。nmf193突变的杂合子小鼠在2个月时表现出较轻的表型,即外段缩短,到10个月时光感受器细胞逐渐丧失50%。采用DNA池策略的定位克隆方法来鉴定nmf193小鼠中的致病突变。nmf193突变与17号染色体连锁,并精细定位到包含外周蛋白/视网膜变性慢(Prph2)基因的区间。突变分析确定Prph2中有一个单碱基变化,导致外显子1和2之间的异常剪接。有趣的是,比较组织学分析表明,Prph2(nmf193/+)突变体与Nr2e3(rd7/rd7)具有相似的光感受器变性。我们发现,与对照同窝小鼠相比,Nr2e3(rd7/rd7)突变体中Prph2 mRNA和蛋白水平降低。染色质免疫沉淀分析表明Prph2是NR2E3的直接靶点。此外,Nr2e3(rd7/rd7)和Prph2(nmf193/+)突变体中Prph2基因表达的下调相似,这表明Prph2的减少可能导致Nr2e3(rd7/rd7)中所见的退行性病理变化。