Analysis of the roles of POMGnT1 in development and maintenance of certral nervous system and skeletal muscle

POMGnT1在中枢神经系统和骨骼肌发育和维持中的作用分析

• 批准号: 18590392
• 负责人: SUZUKI Yuko
• 金额: $ 2.57万
• 依托单位: National Center of Neurology and Psychiatry
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590392/
• 关键词: glycosylation; POMGnT1; alpha-dystroglycan; skeletal muscle; muscle satallite cells; muscle-eye-brain disease; laminin; neuronal migration

Protein O-linked mannose β1, 2-N-acetylglucosaminyltransferase 1(POMGnT1) is an enzyme that transfers N-acetylglucosamine to O-mannose of glycoproteins, and is indispensable for (O-mannosyl glycosylation of alpha-dystroglycan(α-DG), a component of the dystrophin-glycoprotein complex. Mutation of the POMGnT1 gene is a cause of muscle-eye-brain(MEB) disease, which is characterized by severe congenital muscular dystrophy and abnormalities of the eyes and central nervous system(CNS). To obtain a better understanding of the pathogenesis of MEB disease, we mutated the POMGnT1 gene in mice using a targeting technique.1) Skeletal muscle The mutant muscle showed aberrant glycosylation of α-DG and α-DG from mutant muscle failed to bind laminin in a binding assay. POMGnT1^<-/-> muscle showed minimal pathological changes with very low serum creatinine kinase levels, and had normally formed muscle basal lamina. Importantly, POMGnT1^<-/-> satellite cells proliferated slowly, but efficiently differen … More tiated into multinuclear myotubes in vitro. Transfer of a retrovirus vector-mediated POMGnT1 gene into POMGnT1^<-/-> myoblasts completely restored the glycosylation of α-DG, but proliferation of the cells was not improved. Our results suggest that α-DG is not directly involved in the proliferation and differentiation of myoblasts but that its glycosylation is important for maintenance of the proliferative capacity of satellite cells in the postnatal stage.2) CNS The cerebral cortex of POMGnT1^<-/-> mice showed abnormal migration of neurons, disrupted glia limitans and reactive gliosis. The expression of reelin was widely up-regulated in the mutant neocortex. We tested whether the restoration of POMGnT1 expression in neuron results in normal migration along the radial glia. To this end, we electroporated a POMGnT1-expressing plasmid into the ventricular zone of E12 and E15 embryos in utero.Surprisingly, the migration pattern of POMGnT1-deficient neuron did not change after restoration of POMGnT1 expression. The results suggest that the abnormal migration of POMGnT1^<-/-> neurons is not due to deficiency ogf POMGn1 activity in neuron but likely due to abnormality of radial glial cells(Bergmann glia), which have long radial processes extending from the ventricular zone(VZ) to the pial surface and serve a railroad of neuronal migration. Less

蛋白质O-连接甘露糖β1，2-N-乙酰葡糖胺转移酶1（POMGnT 1）是一种将N-乙酰葡糖胺转移至糖蛋白的O-甘露糖的酶，并且是α-肌营养不良蛋白聚糖（α-DG）（肌营养不良蛋白-糖蛋白复合物的组分）的O-甘露糖基化所必需的。POMGnT 1基因突变是肌-眼-脑（MEB）疾病的原因，其特征是严重的先天性肌营养不良症以及眼睛和中枢神经系统（CNS）的异常。为了更好地了解MEB疾病的发病机制，我们使用靶向技术在小鼠中突变POMGnT 1基因。1）骨骼肌突变肌肉显示α-DG的异常糖基化，并且在结合试验中突变肌肉的α-DG不能结合层粘连蛋白。POMGnT 1 ^<-/->肌肉显示出最小的病理变化，血清肌酐激酶水平非常低，并且具有正常形成的肌肉基底层。重要的是，POMGnT 1 ^<-/->卫星细胞增殖缓慢，但能有效地抑制肿瘤细胞的增殖。 ...更多信息 体外诱导成多核肌管。将逆转录病毒载体介导的POMGnT 1基因转移到POMGnT 1 ^<-/->成肌细胞中，可完全恢复α-DG的糖基化，但细胞增殖没有改善。结果表明，α-DG不直接参与成肌细胞的增殖和分化，但其糖基化对维持出生后阶段卫星细胞的增殖能力具有重要作用。2）中枢神经系统POMGnT 1 ^<-/->小鼠大脑皮层神经元迁移异常，胶质细胞界膜破坏，反应性胶质增生。reelin的表达在突变的新皮层中广泛上调。我们测试了神经元中POMGnT 1表达的恢复是否导致沿放射状胶质沿着的正常迁移。为此，我们将表达POMGnT 1的质粒电穿孔到子宫内E12和E15胚胎的脑室区。令人惊讶的是，POMGnT 1缺陷神经元的迁移模式在恢复POMGnT 1表达后没有改变。结果提示，POMGn T 1 ^<-/->神经元的异常迁移不是由于神经元中POMGn 1活性的缺乏，而可能是由于放射状胶质细胞（Bergmann glia）的异常所致。放射状胶质细胞具有从脑室区（VZ）延伸到软膜表面的长放射状突起，为神经元迁移提供了一条铁路。少

项目成果

Molecular signature of quiescent satellite cells in adult skeletal muscle

• DOI: 10.1634/stemcells.2007-0019
• 发表时间: 2007-01-01
• 期刊: STEM CELLS
• 影响因子: 5.2
• 作者: Fukada, So-Ichiro;Uezumi, Akiyoshi;Takedaa, Shin'ichi
• 通讯作者: Takedaa, Shin'ichi

CD90-positive cells, an additional cell population, produce laminin <alpha>2 upon transplantation to dy3k/dy3k mice.

CD90阳性细胞，一种额外的细胞群，在移植到dy3k/dy3k小鼠后产生层粘连蛋白α2。

• 发表时间: 2008
• 期刊: Exp.Cell Res. 314
• 作者: Fukada S;Yamamoto Y;Segawa M;Sakamoto K;Nakajima M;Sato M;Morikawa D;Uezumi A;Miyagoe-Suzuki Y;Takeda S;Tsujikawa K;Yamamoto H
• 通讯作者: Yamamoto H

CD90-positive cells.an additional cell population, produce laminin alpha2 upon transplantation to dy(3k)/dyd3k)mice

CD90 阳性细胞。额外的细胞群，移植到 dy(3k)/dyd3k) 小鼠后产生层粘连蛋白 α2

• 发表时间: 2008
• 期刊: Experimental Cell Research 314
• 作者: Fukada S;Yamamoto Y;Segawa M;Sakamoto K;Nakajima M;Sato M;Morikawa D;Uezumi A;Miyagoe-Suzuki Y;Takeda S;Tsuiikawa K;Yamamoto H.
• 通讯作者: Yamamoto H.

Co-transplantation of muscle-derived side population(SP)cells with myoblasts promoted muscle regeneration

肌源侧群（SP）细胞与成肌细胞共移植促进肌肉再生

• 发表时间: 2007
• 作者: Norio Motohashi;Akiyoshi Uezumi;Erica Yada;So-ichirou Fukada;Kazuhiko Imaizumi;Yuko Miyagoe-Suzuki;Shin'ichi Takada
• 通讯作者: Shin'ichi Takada

Autologous transplantation of SM/C-2.6+ satellite cells transduced with micro-dystrophin CS1 cDNA by lentiviral vector into mdx mice

• DOI: 10.1038/sj.mt.6300295
• 发表时间: 2007-12-01
• 期刊: MOLECULAR THERAPY
• 影响因子: 12.4
• 作者: Ikemoto, Madoka;Fukada, So-ichiro;Takeda, Shin'ichi
• 通讯作者: Takeda, Shin'ichi