Holzfeind Paul J, Grewal Prabhjit K, Reitsamer Herbert A, Kechvar Jasmin, Lassmann Hans, Hoeger Harald, Hewitt Jane E, Bittner Reginald E
Neuromuscular Research Department, Institute of Anatomy, University of Vienna Medical School, 1090 Vienna, Austria.
Hum Mol Genet. 2002 Oct 1;11(21):2673-87. doi: 10.1093/hmg/11.21.2673.
We have recently shown that a deletion in the Large gene, encoding a putative glycosyltransferase, is the molecular defect underlying the myodystrophy (previously myd; now Large(myd)) mouse. Here we show that the muscular dystrophy phenotype is not confined to skeletal muscle, but is also present in the heart and tongue. Immunohistochemistry indicates disruption of the dystrophin-associated glycoprotein complex (DGC) in skeletal and cardiac muscle. Quantitative western blotting shows a general increase in the expression of DGC proteins and of dysferlin and caveolin-3 in mutant skeletal muscle. In contrast, the expression of DGC proteins is reduced in cardiac muscle. Overlay assays show loss of laminin binding by alpha-dystroglycan in Large(myd) skeletal and cardiac muscle and in brain. We also show that the phenotype of Large(myd) mice is not restricted to muscular dystrophy, but also includes ophthalmic and central nervous system (CNS) defects. Electroretinograms of homozygous mutant mice show gross abnormalities of b-wave characteristics, indicative of a complex defect in retinal transmission. The laminar architecture of the cortices of the cerebrum and the cerebellum is disturbed, indicating defective neuronal migration. Thus, the phenotype of the Large(myd) mouse shows similarities to the heterogeneous group of human muscle eye brain diseases characterized by severe congenital muscular dystrophy, eye abnormalities and CNS neuronal migration defects. These diseases include Fukuyama-type muscular dystrophy and muscle-eye-brain disease, both of which are also due to mutations in predicted glycosylation enzymes. Therefore, the Large(myd) mouse represents an important animal model for studying the function of glycosylation in muscle, brain and retina.
我们最近发现,编码一种假定糖基转移酶的Large基因中的一个缺失,是肌营养不良症(以前称为myd;现在称为Large(myd))小鼠的分子缺陷所在。在此我们表明,肌肉萎缩表型不仅局限于骨骼肌,在心脏和舌头中也存在。免疫组织化学表明,骨骼肌和心肌中肌营养不良蛋白相关糖蛋白复合体(DGC)遭到破坏。定量蛋白质免疫印迹分析显示,突变骨骼肌中DGC蛋白、dysferlin和小窝蛋白-3的表达普遍增加。相比之下,心肌中DGC蛋白的表达减少。覆盖分析显示,在Large(myd)小鼠的骨骼肌、心肌和脑中,α- dystroglycan与层粘连蛋白的结合丧失。我们还表明,Large(myd)小鼠的表型不仅限于肌肉萎缩,还包括眼部和中枢神经系统(CNS)缺陷。纯合突变小鼠的视网膜电图显示b波特征存在明显异常,表明视网膜传导存在复杂缺陷。大脑和小脑皮质的分层结构受到干扰,表明神经元迁移存在缺陷。因此,Large(myd)小鼠的表型与人类肌肉-眼-脑疾病的异质性群体相似,这些疾病的特征是严重的先天性肌肉萎缩、眼部异常和CNS神经元迁移缺陷。这些疾病包括福山型肌肉萎缩症和肌肉-眼-脑疾病,两者也都是由于预测的糖基化酶发生突变所致。因此,Large(myd)小鼠是研究糖基化在肌肉、大脑和视网膜中功能的重要动物模型。