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O-连接的甘露糖β1，2-N-乙酰葡萄糖胺1（POMGNT1）是一种将N-乙酰基葡萄糖胺转移到糖蛋白的O-甘露糖中的酶，对于O-甘露糖基糖基化的alpha-dgglylycun（o-甘露糖基）是不可见的pomgnt1基因的肌蛋白糖蛋白复合物是肌肉 - 脑（MEB）疾病的原因，其特征是严重的先天性肌肉肌营养不良，眼睛和中枢神经系统的异常（CNS）可以更好地理解MEB疾病。肌肉突变肌显示出来自突变肌肉的α-DG和α-DG的异常糖基化未能结合结合测定中的层粘连蛋白。 pomgnt1^< - / - >肌肉显示出最小的病理变化，血清肌酐激酶水平非常低，并且通常形成肌肉碱性薄片。重要的是，pomgnt1^< - / - >卫星细胞在体外缓慢增殖，但有效地不同……在体外更加融入多核肌管。将逆转录病毒介导的pomgnt1基因转移到Pomgnt1^< - / - >成肌细胞中完全恢复了α-DG的糖基化，但是细胞的增殖没有得到改善。 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 proliferation capacity of satellite cells in the postnatal stage.2) CNS The cerebral cortex of POMGnT1^<-/-> mice shown abnormal migration of neurons, disrupted glia limitans and reactive gliosis. reelin的表达在突变的新皮层中被广泛上调。我们测试了神经元中POMGNT1表达的恢复是否导致沿着径向神经胶质的正常迁移。为此，我们将表达POMGNT1的质粒电穿孔到子宫内E12和E15胚胎的心室区域。令人震惊的是，POMGNT1缺乏性神经元的迁移模式在POMGNT1表达恢复后没有变化。结果表明，pomgnt1^< - / - >神经元的异常迁移不是由于神经元中的OGF POMGN1活性不足，但可能是由于径向神经胶质细胞（Bergmann Glia）的异常引起的，这些异常具有长时间的辐射过程，这些过程长期从心室（VZ）延伸至pie Surface and pie Surface and a Rail of neuronal迁移。较少的

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 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

Injection of a recombinant AAV serotype 2 into canine skeletal muscles evokes strong immune responses against transgene products

• DOI: 10.1038/sj.gt.3302984
• 发表时间: 2007-09-01
• 期刊: GENE THERAPY
• 影响因子: 5.1
• 作者: Yuasa, K.;Yoshimura, M.;Takeda, S.
• 通讯作者: Takeda, S.

Downstream utrophin enhancer is required for expression of utrophin in skeletal muscle.

骨骼肌中肌营养蛋白的表达需要下游肌营养蛋白增强剂。

• 发表时间: 2008
• 期刊: Journal of Gene Medicine 10
• 作者: Tanihata J;Suzuki N;Miyagoe-Suzuki Y;Imaizumi K;Takeda S.
• 通讯作者: Takeda S.