Dhaenens C M, Schraen-Maschke S, Tran H, Vingtdeux V, Ghanem D, Leroy O, Delplanque J, Vanbrussel E, Delacourte A, Vermersch P, Maurage C A, Gruffat H, Sergeant A, Mahadevan M S, Ishiura S, Buée L, Cooper T A, Caillet-Boudin M L, Charlet-Berguerand N, Sablonnière B, Sergeant N
Inserm, U837, place de Verdun, 59045 Lille, France.
Exp Neurol. 2008 Apr;210(2):467-78. doi: 10.1016/j.expneurol.2007.11.020. Epub 2007 Dec 4.
Neurofibrillary degeneration is often observed in the brain of patients with type 1 myotonic dystrophy (DM1). It consists principally of the aggregation of Tau isoforms that lack exon 2/3 encoded sequences, and is the consequence of the modified splicing of Tau pre-mRNA. In experimental models of DM1, the splicing of several transcripts is modified due to the loss of Muscleblind-like 1 (MBNL1) function. In the present study, we demonstrate that the MBNL1 protein is also present in the human brain, and consists of several isoforms, as shown by RT-PCR and sequencing. In comparison with controls, we show that the adult DM1 brain exhibits modifications in the splicing of MBNL1, with the preferential expression of long MBNL1 isoforms--a splicing pattern similar to that seen in the fetal human brain. In cultured HeLa cells, the presence of long CUG repeats, such as those found in the DM1 mutation, leads to similar changes in the splicing pattern of MBNL1, and the localization of MBNL1 in nuclear RNA foci. Long CUG repeats also reproduce the repression of Tau exon 2/3 inclusion, as in the human disease, suggesting that their effect on MBNL1 expression may lead to changes in Tau splicing. However, while an overall reduction in the expression of MBNL1 mimics the effect of the DM1 mutation, none of the MBNL1 isoforms tested so far modulates the endogenous splicing of Tau. The modified splicing of Tau thus results from a possibly CUG-mediated loss of function of MBNL1, but not from changes in the MBNL1 expression pattern.
在1型强直性肌营养不良(DM1)患者的大脑中经常观察到神经原纤维变性。它主要由缺乏外显子2/3编码序列的Tau异构体聚集而成,是Tau前体mRNA剪接改变的结果。在DM1的实验模型中,由于类肌肉盲蛋白1(MBNL1)功能丧失,几种转录本的剪接发生改变。在本研究中,我们证明MBNL1蛋白也存在于人类大脑中,并且由几种异构体组成,如逆转录聚合酶链反应(RT-PCR)和测序所示。与对照组相比,我们发现成年DM1患者大脑中MBNL1的剪接存在改变,长MBNL1异构体优先表达——这种剪接模式与人类胎儿大脑中的相似。在培养的HeLa细胞中,长CUG重复序列(如在DM1突变中发现的那些)的存在导致MBNL1剪接模式发生类似变化,以及MBNL1在核RNA灶中的定位。长CUG重复序列也像在人类疾病中一样再现了对Tau外显子2/3包含的抑制,这表明它们对MBNL1表达的影响可能导致Tau剪接的变化。然而,虽然MBNL1表达的总体降低模拟了DM1突变的作用,但到目前为止测试的任何一种MBNL1异构体都没有调节Tau的内源性剪接。因此,Tau剪接的改变可能是由CUG介导的MBNL1功能丧失引起的,而不是由MBNL1表达模式的变化引起的。