TOMOGRAPHY OF SKELETAL MUSCLE TRIADIC JUNCTION

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 摘要： 我们正在将我们对骨骼肌中ryanodine受体的研究扩展到原位观察，希望能够可视化二氢吡啶受体（介导电脉冲和引起收缩的钙释放的电压传感器）。 到目前为止，在Wagenknecht博士的实验室中，在单颗粒兰尼碱受体的工作和孤立三联体连接的工作中，二氢吡啶受体在分离过程中要么被去除，要么被破坏。 我们正在与宾夕法尼亚大学的克拉拉·弗兰兹尼-阿姆斯特朗博士合作，她是兴奋-收缩装置研究的领导者。 受益于她在准备肌肉组织的最佳可视化的三联体连接在原位的长期经验，我们正在开发的方法，高压冷冻肌肉组织在适当的方向观察的二氢吡啶受体。 在上一个报告所述期间，Franzini-Armstrong博士访问了资源中心，并在一次研讨会上介绍了该项目。 她还与Chyongere Hsieh合作准备高压冷冻标本。 断层重建是由Franzini-Armstrong博士提供的薄的常规制备的塑料切片制成的。 这样做是为了学习识别肌肉纤维的理想方向，以揭示DHPR。

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.