Search for aquaporin molecules at the plasma membrane of normal skeletal myofibers and their alterations in myopathic muscles.

寻找正常骨骼肌纤维质膜上的水通道蛋白分子及其在肌病肌肉中的变化。

• 批准号: 14570620
• 负责人: WAKAYAMA Yoshihiro
• 金额: $ 2.24万
• 依托单位: Showa 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 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570620/
• 关键词: Aquaporins; Skeletal myofiber; Normal and disease; RNA; Protein; RT-PCR; Immunoblot; Immunohistochemistry; 正常骨格筋; アクアポリン3; アクアポリン4; アクアポリン7; mRNA; 免疫組織化学; 福山型筋ジストロフィー症生検筋; アクアポリン; 骨格筋; 免疫電顕

We investigated what kind of aquaporin molecule is expressed in normal skeletal muscle and the investigation was extended to analyze the alteration of the expression of these aquaporin molecule in the diseased human muscles. With regard to the expression of aquaporin molecule in normal skeletal muscle, each of aquaporin 0〜10 molecule was searched at RNA and protein levels. The investigation of RNA level was carried out by RT PCR and that of protein level was performed by immunoblot and immunocytochemistry As the result, AQP3, AQP4 AQP7 molecules were confirmed to be present at RNA and protein levels in normal human skeletal myofibers. AQP4 molecule was more abundantly expressed in type 2 white fiber (fast twitch fiber) than in type 1 red fiber (slow twitch fiber): whereas AQP3 molecule was more abundantly expressed in type 1 red fiber than in type 2 white fiber. AQP7 molecule was equally expressed in type 1 and type 2 human skeletal myofibers; while in normal mouse skeletal muscle AQP7 … More was more abundantly expressed in type 2 fast twitch fiber than type 1 slow twitch fiber. AQP 1 is reported to be present at the endothelial cell within skeletal muscle tissue, however; we noticed that AQP 1 is present at the myofiber surface of skeletal muscle tissue. Electron microscopy of fracture label replica of normal skeletal muscle showed that gold signal of AQP4 was located in the orthogonal array of myofiber P face and that of AQP3 was located in the particle cluster of myofiber P face. Particle cluster is the irregular aggregate of intramembranous particles and is different from orthogonal array which is the regular particle aggregate more than 4 particles forming rectangular shape. With respect to the altered expression of aquaporin molecule in diseased human skeletal muscles, expression of AQP4 in muscles with Fukuyama type congenital muscular dystrophy was reduced in their myofiber surface at protein levels and the content of AQP4 mRNA was reduced AQP4 expression was also decreased in the scattered myofibers with neurogenic atrophy muscles such as muscles with amyotrophic lateral sclerosis. Less

我们研究了什么样的水通道蛋白分子在正常骨骼肌中表达，并将研究扩展到分析这些水通道蛋白分子在患病人体肌肉中表达的变化。关于水通道蛋白分子在正常骨骼肌中的表达，在RNA和蛋白质水平上搜索水通道蛋白0 - 10分子中的每一个。用RT PCR法检测了正常人骨骼肌纤维中的RNA水平，用免疫印迹法和免疫细胞化学法检测了蛋白水平。结果表明，正常人骨骼肌纤维中存在AQP 3、AQP 4和AQP 7分子。AQP 4分子在2型白色纤维（快缩纤维）中的表达高于1型红纤维（慢缩纤维），而AQP 3分子在1型红纤维中的表达高于2型白色纤维。AQP 7分子在人1型和2型骨骼肌纤维中表达相同，而在正常小鼠骨骼肌AQP 7 ...更多信息 在2型快缩纤维中的表达较1型慢缩纤维丰富。据报道，AQP 1存在于骨骼肌组织内的内皮细胞，然而，我们注意到，AQP 1存在于骨骼肌组织的肌纤维表面。正常骨骼肌断裂标记复制品的电镜观察显示，AQP 4的金信号位于肌纤维P面的正交排列中，AQP 3的金信号位于肌纤维P面的颗粒簇中。颗粒簇是膜内颗粒的不规则聚集体，不同于正交排列，正交排列是4个以上的颗粒聚集成矩形。关于水通道蛋白分子在患病的人骨骼肌中的表达改变，在Fukuyama型先天性肌营养不良症的肌肉中，AQP 4在其肌纤维表面在蛋白水平上的表达降低，并且AQP 4 mRNA的含量降低。在神经源性萎缩的肌肉中，如肌萎缩侧索硬化症的肌肉中，AQP 4表达也降低。少

项目成果

Expression of aquaporin 3 and its localization in normal skeletal myofibres.

水通道蛋白 3 的表达及其在正常骨骼肌纤维中的定位。

• 发表时间: 2002
• 期刊: Histochem J 34(6-7)
• 作者: Wakayama Y;Jimi T;Inoue M;et al.;Sasaki S et al.;Sonoo M.;Shibata N et al.;Hatanaka Y et al.;Yoshihiro Wakayama
• 通讯作者: Yoshihiro Wakayama

Takahiro Jimi: "Reduced expression of aquaporin 4 in human muscles with amyotrophic lateral sclerosis and other neurogenic atrophies."Pathology Reserarch and Practice. 印刷中.

Takahiro Jimi：“患有肌萎缩侧索硬化症和其他神经源性萎缩的人体肌肉中水通道蛋白 4 的表达减少。”病理学研究与实践。

Yoshihiro Wakayama: "Expression and localization of aquaporin 7 in normal skeletal myofiber."Cell and Tissue Research. 印刷中.

Yoshihiro Wakayama：“水通道蛋白 7 在正常骨骼肌纤维中的表达和定位。”细胞和组织研究正在出版。

Expression and localization of aquaporin 7 in normal skeletal myofiber.

水通道蛋白 7 在正常骨骼肌纤维中的表达和定位。

• 发表时间: 2004
• 期刊: Cell Tissue Res 316(1)
• 作者: Sonoo M;Tsaiweichao-Shozawa Y;Oshimi-Sekiguchi M et al.;Takahiro Jimi;Sonoo M et al.;Yoshihiro Wakayama
• 通讯作者: Yoshihiro Wakayama

Yoshihiro Wakayama: "Altered aquaporin 4 expression in muscles of Fukuyama-type congenial muscular dystrophy."Virchows Archiv. 443(6). 761-767 (2003)

Yoshihiro Wakayama：“改变了福山型先天性肌营养不良症肌肉中的水通道蛋白 4 表达。”Virchows Archive。