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Research on molecular pathogenesis of congenital muscular dystrophy -Characterization of the intra-/extra-cellular interaction of dystroglycan-

先天性肌营养不良症的分子发病机制研究-肌营养不良症细胞内/外相互作用的表征-

基本信息

批准号：
17590898
负责人：
SAITO Fumiaki
金额：
$ 2.24万
依托单位：
Teikyo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590898/
关键词：
Muscular dystrophy Dystroglycan Glycosylation Fukutin Neuromuscular junction Laminin Agrin 糖鎖髄鞘蛋白質

项目摘要

To clarify the molecular mechanisms underlying congenital muscular dystrophy, I characterized the skeletal muscle, neuromuscular junction and peripheral nerve of dystrophic chicken and fukutin deficient chimeric mouse, animal models of congenital muscular dystrophy. First, analysis using dystrophic chicken revealed that 1) glycosylation of α-dystroglycan is defective and its laminin binding activity is reduced in the skeletal muscle of dystrophic chicken, 2) two distinct molecular species of α-dystroglycan exists in the chicken and the smaller species doesn't possess the laminin binding activity, 3) amount of Galβ1-3GalNAc moiety is increased whereas Siaa2-3Gal is reduced on α-dystroglycan of the dystrophic chicken as compared to control 4) expression of β1-integron is up-regulated in the skeletal muscle and heart of the chicken. Second, experiments with fukutin deficient chimeric mice demonstrated that 1) neuromuscular junction of the chimeric mice is small and fragmented in appearance, 2) agrin binding activity and its expression is reduced in the chimeric neuromuscular junction, 3) the radial sorting mechanism of peripheral nerve axon is defective and myelinated fiber is decreased in the chimeric mice, 4) α-dystroglycan is aberrantly glycosylated and its laminin binding activity is decreased in the chimeric nerve. In addition, by yeast two-hybrid screening of human cDNA library, I cloned plasma membrane Ca^<2+> ATPase as a potential binding protein of β-dystroglycan. However, the molecular interaction of these proteins was not confirmed at protein level with co-immunoprecipitation experiments or pull-down assays, suggesting that the interaction is so weak that these methodology couldn't detect the binding.

为了阐明先天性肌营养不良的分子机制，我对先天性肌营养不良的动物模型--营养不良鸡和Fukutin缺陷嵌合小鼠的骨骼肌、神经肌肉接头和周围神经进行了表征。首先，通过对营养不良鸡的分析发现：1)营养不良鸡骨骼肌中α-dystroglan的糖基化存在缺陷，其层粘连蛋白结合活性降低；2)营养不良鸡中存在两种不同的α-dystroglan分子，较小的种不具有层粘连蛋白结合活性；3)与对照相比，营养不良鸡的β1-3GalNAc部分的含量增加而Sia2-3Gal的含量减少；4)α-整合子在骨骼肌和心脏中的表达上调。2)聚集素结合活性及其表达降低；3)嵌合小鼠周围神经轴突的径向分选机制存在缺陷，有髓纤维减少；4)α-dystroglan糖基化异常，其层粘连蛋白结合活性在嵌合神经中降低。此外，通过酵母双杂交筛选文库，克隆了质膜Ca^&lt；2+&Gt；ATPase作为β-DystroGan的潜在结合蛋白。然而，免疫共沉淀实验或下拉实验没有在蛋白质水平上证实这些蛋白质之间的相互作用，这表明这种相互作用非常弱，以至于这些方法无法检测到这种结合。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Defective peripheral nerve myelination and neuromuscular junction form ation in fukutin-deficient chimeric mice.

fukutin 缺陷嵌合小鼠周围神经髓鞘形成和神经肌肉接头形成缺陷。

DOI：
发表时间：
2007
期刊：
J. Neurochem. 101
影响因子：
0
作者：
Yoshida T;Yazaki M;Gono T;Tazawa K;Morita H;Matsuda M;Funakoshi K;Yuki N;Ikeda S.;Hara-Chikuma M et al.;Saito F
通讯作者：
Saito F

Aberrant glycosylation of α-dystroglycan causes defective binding of laminin in the muscle of chicken muscular dystrophy