Role of perlecan at the neuromuscular junction

基底膜蛋白聚糖在神经肌肉接头中的作用

• 批准号: 14570618
• 负责人: HIRASAWA Eri
• 金额: $ 2.24万
• 依托单位: Juntendo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570618/
• 关键词: perlecan; peparan sulfate proteoglycan; neuromuscular junction; acetylcholine esterase; myotonia; microelectrode analyses; Schwartz-Jampel syndrome

At the NMJ, the nicotinic ACh receptor mediates postsynaptic depolarization, and acetylcholinesterase (AChE) terminates this process by hydrolyzing ACh. Efficient and accurate synaptic transmission requires proper localization of many signaling proteins in the synaptic membrane. In the perlecan knockout mice, muscle development and differentiation appear to be normal and normal nerve terminals are formed at birth. Clustering molecules are present at the NMJ of the mutant mouse muscles. However, AChE is absent at the newborn MNJ, although AChE protein is synthesized normally. An animal model is ideal to study the mechanism of myotonia of SJS(Schwartz-Jampel syndrome). However, perlecan-null mice are lethal. To overcome this problem, we used a genetic approach to rescue the perinatal lethal phenotype of perlecan-null mice by mating heterozygotes of perlecan-null mice with transgenic mice expressing recombinant perlecan specifically to cartilage but not to other tissues. The perlecan-null homozygous mice with the transgene are born normal and survive, but they develop small eyes and blepharospasm, common features of SJS. These mice also show myotonic discharge in electromyography (EMG) and myopathic change in histology. AChE activity staining shows reduced localization of AChE at the NMJ in the mutant. mice. Thus, the rescued mice show phenotypes characteristic of SJS and are useful as an animal model to study the mechanism of myotonia and to develop therapeutic agents for the disease.

在NMJ，烟碱ACh受体介导突触后去极化，乙酰胆碱酯酶（AChE）通过水解ACh终止这一过程。有效和准确的突触传递需要许多信号蛋白在突触膜中的适当定位。在串珠素基因敲除的小鼠中，肌肉发育和分化似乎是正常的，并且在出生时形成了正常的神经末梢。聚集分子存在于突变小鼠肌肉的NMJ处。然而，AChE是缺乏在新生MNJ，虽然AChE蛋白合成正常。动物模型是研究SJS（Schwartz-Jampel综合征）肌强直机制的理想模型。然而，无串珠素蛋白聚糖的小鼠是致命的。为了克服这个问题，我们使用了一种遗传方法来拯救围产期致死表型的串珠素基因敲除小鼠交配杂合子串珠素基因敲除小鼠与转基因小鼠表达重组串珠素特异性软骨，但不对其他组织。带有转基因的串珠蛋白聚糖缺失纯合子小鼠出生正常并存活，但它们发展为小眼睛和眼睑痉挛，这是SJS的共同特征。这些小鼠还在肌电图（EMG）中显示肌强直放电和在组织学中的肌病改变。AChE活性染色显示突变体中AChE在NMJ处的定位减少。小鼠因此，获救的小鼠显示出SJS的表型特征，并且可用作研究肌强直机制和开发该疾病的治疗剂的动物模型。

项目成果

Hirasawa M, Ohshima T, Takahashi S, et al.: "Perinatal abrogation of Cdk5 expression in brain results in neuronal migration defects"Proc Natl Acad Sci U S A.. 101(16). 6249-6254 (2004)

Hirasawa M、Ohshima T、Takahashi S 等人：“脑中 Cdk5 表达的围产期废除导致神经元迁移缺陷”Proc Natl Acad Sci U S A.. 101(16)。

Arikawa-Hirasawa E, Yamada Y: "Perlecan mutations in mice and humans : critical role of perlecanin skeletal development and disease"Acta myologica. XXSeptember. 134-137 (2002)

Arikawa-Hirasawa E、Yamada Y：“小鼠和人类的基底膜蛋白突变：基底膜蛋白骨骼发育和疾病的关键作用”《肌学报》。

Yuasa K, Fukumoto S, Kamasaki Y, Yamada A, Fukumoto E, Kanaoka K, Saito K, Harada H, Arikawa-Hirasawa E, Miyagoe-Suzuki Y, Takeda S, Okamoto K, Kato Y, Fujiwara T.: "Laminin alpha2 essential for odontoblast differentiation regulating dentin sialoprotein e

汤浅 K、福本 S、镰崎 Y、山田 A、福本 E、金冈 K、斋藤 K、原田 H、有川平泽 E、宫越铃木 Y、武田 S、冈本 K、加藤 Y、藤原 T.：“层粘连蛋白 α2

Arikawa-Hirasawa E, Wilcox WR, Yamada Y: "Dyssegmental Dysplasia, Silverman-Handmaker Type : Unexpected role of perlecan in cartilage development."Am.J.Med.Gen.. 106. 254-257 (2002)

Arikawa-Hirasawa E、Wilcox WR、Yamada Y：“节段性发育不良，Silverman-Handmaker 类型：基底膜蛋白在软骨发育中的意外作用。”Am.J.Med.Gen.. 106. 254-257 (2002)

Yuasa K, Fukumoto S, Kamasaki Y, et al.: "Laminin alpha2 essential for odontoblast differentiation regulating dentin sialoprotein expression"J Biol Chem.. 279. 10286-10292 (2003)

Yuasa K、Fukumoto S、Kamasaki Y 等：“成牙本质细胞分化所必需的层粘连蛋白 α2 调节牙本质唾液蛋白表达”J Biol Chem.. 279. 10286-10292 (2003)