TOMOGRAPHY OF SKELETAL MUSCLE TRIADIC JUNCTION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 摘要： 我们正在将我们对骨骼肌中ryanodine受体的工作扩展到原位观察，希望可视化二氢吡啶受体(介导电脉冲和导致收缩的钙释放的电压传感器)。到目前为止，在瓦根克内赫特博士的实验室中，在单粒子Ryanodine受体的工作以及对孤立的三联体连接的工作中，二氢吡啶受体要么在分离过程中被移除，要么被破坏。我们正在与宾夕法尼亚大学的克拉拉·弗兰齐尼-阿姆斯特朗博士合作，他是兴奋收缩装置研究的领先者。凭借她在制备肌肉组织以优化原位三联体连接可视化方面的长期经验，我们正在开发以适当方向高压冷冻肌肉组织以观察二氢吡啶受体的方法。 在上一个报告所述期间，弗兰齐尼-阿姆斯特朗博士访问了资源中心，并举办了一次研讨会，介绍了该项目。她还与谢宗怡合作，为高压冷冻准备标本。断层重建是由弗兰齐尼-阿姆斯特朗博士提供的薄薄的、传统准备的塑料切片制成的。这样做是为了学习识别揭示DHPR的肌肉纤维的理想方向。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. ABSTRACT: We are extending our work on the ryanodine receptor in skeletal muscle to in-situ observation, with the hope of visualizing dihydropyridine receptors (the voltage sensors that mediate the electrical impulse and the release of calcium that causes contraction). In the single-particle ryanodine receptor work, and the work on isolated triad junctions, done so far in Dr. Wagenknecht's lab, the dihydropyridine receptors were either removed or disrupted during isolation. We are collaborating with Dr. Clara Franzini-Armstrong of the University of Pennsylvania, who is a leader in the study of the excitation-contraction apparatus. With the benefit of her long experience in preparing muscle tissue for optimal visualization of the triad junction in situ, we are developing methods for high-pressure freezing muscle tissue in the proper orientation for observation of the dihydropyridine receptors. In the previous reporting period, Dr. Franzini-Armstrong visited the Resource and presented a seminar to describe the project. She also worked with Chyongere Hsieh in preparing specimens for high-pressure freezing. Tomographic reconstructions were made from thin, conventionally-prepared plastic sections provided by Dr. Franzini-Armstrong. This was done in order to learn to recognize the ideal orientation of the muscle fiber for revealing the DHPR.