Muscleblind-like 1 knockout mice reveal novel splicing defects in the myotonic dystrophy brain.
Muscleblind-like 1 knockout mice reveal novel splicing defects in the myotonic dystrophy brain.
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DOI：
10.1371/journal.pone.0033218
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Kimura T
中科院分区：
文献类型：
--
作者：
Suenaga K;Lee KY;Nakamori M;Tatsumi Y;Takahashi MP;Fujimura H;Jinnai K;Yoshikawa H;Du H;Ares M Jr;Swanson MS;Kimura T
强直性肌营养不良1型(DM1)是由DMPK基因CTG三核苷酸重复序列扩增(CTGexp)引起的一种多系统疾病。在骨骼肌中，选择性剪接因子类肌球蛋白1(MBNL1)的核隔离解释了在表达致病范围CTGexp的HSA LR转基因小鼠模型中观察到的大多数选择性剪接缺陷。在本研究中，我们讨论了CUGexp RNA隔离MBNL1也导致哺乳动物大脑剪接缺陷的可能性。我们使用剪接敏感的微阵列检测了纯合子MBNL1ΔE3/ΔE3基因敲除小鼠大脑中的核糖核酸。我们使用RT-PCR来验证通过微阵列分析从MBNL1ΔE3/ΔE3基因敲除小鼠和死后DM1患者的脑组织中确定的可选盒式外显子的子集。令人惊讶的是，对MBNL1ΔE3/ΔE3大脑的剪接敏感微阵列分析只发现了14个错误剪接的外显子候选。虽然我们证实其中几个剪接事件在MBNL1基因敲除和DM1大脑中都受到干扰，但在小鼠模型中剪接错误调节的程度明显低于在DM1中观察到的程度。此外，几个备选外显子，包括Grin1外显子4，App外显子7和MAPT外显子3和9，以前曾被报道在人DM1脑中异常剪接，在MBNL1基因敲除脑中正常剪接。CUGexp RNA对MBNL1的隔离导致了DM1脑中的一些异常剪接事件。然而，我们得出结论，其他因素，可能是其他MBNL蛋白，可能有助于DM1大脑中的剪接错误调节。
Myotonic dystrophy type 1 (DM1) is a multi-systemic disorder caused by a CTG trinucleotide repeat expansion (CTGexp) in the DMPK gene. In skeletal muscle, nuclear sequestration of the alternative splicing factor muscleblind-like 1 (MBNL1) explains the majority of the alternative splicing defects observed in the HSA LR transgenic mouse model which expresses a pathogenic range CTGexp. In the present study, we addressed the possibility that MBNL1 sequestration by CUGexp RNA also contributes to splicing defects in the mammalian brain. We examined RNA from the brains of homozygous Mbnl1 ΔE3/ΔE3 knockout mice using splicing-sensitive microarrays. We used RT-PCR to validate a subset of alternative cassette exons identified by microarray analysis with brain tissues from Mbnl1 ΔE3/ΔE3 knockout mice and post-mortem DM1 patients. Surprisingly, splicing-sensitive microarray analysis of Mbnl1 ΔE3/ΔE3 brains yielded only 14 candidates for mis-spliced exons. While we confirmed that several of these splicing events are perturbed in both Mbnl1 knockout and DM1 brains, the extent of splicing mis-regulation in the mouse model was significantly less than observed in DM1. Additionally, several alternative exons, including Grin1 exon 4, App exon 7 and Mapt exons 3 and 9, which have previously been reported to be aberrantly spliced in human DM1 brain, were spliced normally in the Mbnl1 knockout brain. The sequestration of MBNL1 by CUGexp RNA results in some of the aberrant splicing events in the DM1 brain. However, we conclude that other factors, possibly other MBNL proteins, likely contribute to splicing mis-regulation in the DM1 brain.
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通讯作者：
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