Three Gene-Targeted Mouse Models of RNA Splicing Factor RP Show Late-Onset RPE and Retinal Degeneration
Three Gene-Targeted Mouse Models of RNA Splicing Factor RP Show Late-Onset RPE and Retinal Degeneration
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DOI：
10.1167/iovs.10-5194
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发表时间：
2011-01-01
影响因子：
4.4
通讯作者：
Pierce, Eric A.
中科院分区：
文献类型：
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作者：
Graziotto, John J.;Farkas, Michael H.;Pierce, Eric A.
目的.产生参与mRNA剪接的蛋白质的基因突变，包括前mRNA加工因子3，8和31（PRPF 3，8和31），RP 9和SNRNP 200是晚发型遗传性致盲性疾病视网膜色素变性（RP）的常见原因。目前尚不清楚这些普遍表达的基因突变如何导致视网膜特异性疾病。为了研究RP的RNA剪接因子形式的发病机制，作者产生并表征了Prpf 3-T494 M、Prpf 8-H2309 P敲入小鼠的视网膜表型。同时观察Prpf 31基因敲除小鼠视网膜的超微结构。敲入小鼠在其内源性Prpf 3和Prpf 8基因中具有单密码子改变，其模拟人类PRPF 3和PRPF 8中最常见的致病突变。Prpf 31敲除小鼠模拟了PRPF 31患者中鉴定的大多数突变导致的无效等位基因。通过视网膜电图（ERG）、光学和电子显微镜评价基因靶向小鼠的视网膜表型。Prpf 3（+/T494 M）和Prpf 8（+/H2309 P）基因敲入小鼠的RPE细胞在2岁时表现出基础内折叠的丢失和空泡化，在RPE和Bruch[B]膜之间积累无定形沉积物。这些变化在纯合子小鼠中更严重，并且与Prpf 3-T494 M小鼠中的视杆功能降低相关。在一岁的Prpf 31（+/-）小鼠中检测到RPE的类似退行性变化。在所有三种小鼠模型的RPE细胞中发现类似的退行性变化，表明RPE可能是RP的RNA剪接因子形式中受影响的主要细胞类型。在这些小鼠中观察到的相对晚发型表型与RP患者的典型成年发病一致。（Invest Ophthalmol维斯科学。2011; 52：190-198）DOI：10.1167/iovs.10-5194
PURPOSE. Mutations in genes that produce proteins involved in mRNA splicing, including pre-mRNA processing factors 3, 8, and 31 (PRPF3, 8, and 31), RP9, and SNRNP200 are common causes of the late-onset inherited blinding disorder retinitis pigmentosa (RP). It is not known how mutations in these ubiquitously expressed genes lead to retina-specific disease. To investigate the pathogenesis of the RNA splicing factor forms of RP, the authors generated and characterized the retinal phenotypes of Prpf3-T494M, Prpf8-H2309P knockin mice. The retinal ultrastructure of Prpf31-knockout mice was also investigated.METHODS. The knockin mice have single codon alterations in their endogenous Prpf3 and Prpf8 genes that mimic the most common disease causing mutations in human PRPF3 and PRPF8. The Prpf31-knockout mice mimic the null alleles that result from the majority of mutations identified in PRPF31 patients. The retinal phenotypes of the gene targeted mice were evaluated by electroretinography (ERG), light, and electron microscopy.RESULTS. The RPE cells of heterozygous Prpf3(+/T494M) and Prpf8(+/ H2309P) knockin mice exhibited loss of the basal infoldings and vacuolization, with accumulation of amorphous deposits between the RPE and Bruch[b]'s membrane at age two years. These changes were more severe in the homozygous mice, and were associated with decreased rod function in the Prpf3-T494M mice. Similar degenerative changes in the RPE were detected in Prpf31(+/-) mice at one year of age.CONCLUSIONS. The finding of similar degenerative changes in RPE cells of all three mouse models suggests that the RPE may be the primary cell type affected in the RNA splicing factor forms of RP. The relatively late-onset phenotype observed in these mice is consistent with the typical adult onset of disease in patients with RP. (Invest Ophthalmol Vis Sci. 2011; 52: 190-198) DOI:10.1167/iovs.10-5194