KINETICS OF AXONAL PROTEIN SYNTHESIS AND RNA TRANSPORT

这个子项目是许多研究子项目中的一个 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来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 该项目的目的是确定神经细胞的远端过程如何在损伤后调节其蛋白质水平。 这些“轴突过程”是大脑和脊髓中长距离通信所必需的。 如果这些过程被破坏，沟通就会停止，大脑和脊髓的功能就会丧失。 改善受损轴突的再生将恢复大脑和脊柱的功能。 一些证据表明，远端轴突过程可以自主调节再生所需的蛋白质的水平，通过调节这些蛋白质的合成和降解。 直到最近，我们还没有办法解剖再生轴突的蛋白质组，因为可用于研究的材料非常有限，而且经常被其他细胞成分污染。 我们将利用我们实验室开发的轴突制备和UCSF质谱设备的高灵敏度蛋白质组学应用，以确定轴突合成蛋白质的前体如何被靶向运输到轴突中，以及这些前体编码的蛋白质产物的变化。 我们将使用mRNA：蛋白质复合物的亲和纯化来鉴定将mRNA前体转运到轴突中所需的蛋白质。将这些数据与我们实验室正在进行的轴突mRNA分析相结合，将提供远端轴突蛋白质动力学的系统视图。 最终，这些研究将为我们提供一个独特的轴突生物学视角，这是迄今为止尚不可行的，应该会导致大脑和脊髓创伤性损伤后加速再生的新策略。

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. The objective of this project is to determine how the distal processes of nerve cells regulate their protein levels after injury. These 'axonal processes' are needed for long-range communication in the brain and spinal cord. If these processes are disrupted, communication ceases and function of the brain and spinal cord is lost. Improving regeneration of injured axonal processes will restore function to the brain and spinal column. Several lines of evidence indicate that distal axonal processes can autonomously regulate levels of proteins needed for regeneration through modulating synthesis and degradation of these proteins locally. Until recently we have had no means to dissect the proteome of regenerating axons, since the materials available for study are exceptionally limiting and most often contaminated with other cellular constituents. We will take advantage of an axonal preparation that our laboratory has developed and the high sensitivity proteomics applications of the UCSF Mass Spectrometry Facility to determine how the precursors of axonally synthesized proteins are targeted for transport into axons and what becomes of the protein products encoded by these precursors. We will use affinity purification of mRNA: protein complexes to identify the proteins needed for transport of the mRNA precursors into axons. Integrating these data with ongoing axonal mRNA profiling from our laboratory will provide a systematic view of protein dynamics of distal axons. Ultimately, these studies will provide us with a unique perspective of axonal biology that has not been feasible until now and should lead to novel strategies for accelerating regeneration after traumatic injury of the brain and spinal cord.