Ultracentrifugation for the biochemical characterization of biomarkers involved in neurobiological functions

定量评估不同神经元隔间中的突触和信号蛋白构成了七名申请者研究计划的重要组成部分。Western blotting(WB)是实现这一目的必不可少的技术，我们和道格拉斯研究所的其他研究人员也经常使用WB。然而，为了清楚地定位参与突触信号传递的特定蛋白的功能意义，确定它们的精确亚细胞定位是至关重要的。由于道格拉斯研究所现有的所有离心机(&lt；40,000克)的速度都很低，目前不可能用差速离心法分离细胞制剂中的亚细胞细胞器部分。缺乏超离心机给我们的研究进展带来了巨大的挑战。在这里，我们提议申请这一关键设备：Beckman Optima Max-XP桌面超级离心机，用于七名研究人员的研究项目，以解决神经发育、神经免疫学、神经内分泌学以及神经和精神疾病(如慢性疼痛、阿尔茨海默病、严重抑郁症和精神分裂症)的神经生物学方面的重要研究问题。超速离心将使我们能够获得亚细胞细胞器的几个部分，并检查每个部分中感兴趣的蛋白质，从而深入了解其潜在的机制。所要求的设备是完成七名申请者研究计划的无价工具，将对道格拉斯研究所的研究进展产生重大影响。超速离心机是任何尖端分子神经科学研究环境的必备工具，将增强道格拉斯研究人员在国内和国际上的竞争力。

Quantitative evaluation of synaptic and signaling proteins in various neuronal compartments forms an essential part of the research programs of the seven applicants. Western blotting (WB) is an indispensable technique for this purpose and is used routinely by us and other investigators at the Douglas Institute as well. However, in order to clearly pinpoint the functional significance of specific proteins involved in synaptic signalling, it is essential to determine their precise subcellular localization. Differential centrifugation to separate the subcellular organelle fractions of cell preparations is currently impossible due to the low speed of all existing centrifuges (<40,000g) at the Douglas Institute. The absence of an ultracentrifuge imposes a significant challenge to our research progress. Here we are proposing to apply for this essential piece of equipment: Beckman Optima MAX-XP Tabletop Ultracentrifuge for the research programs of seven investigators, to address important research questions in neurodevelopment, neuroimmunology, neuroendocrinology, and neurobiology of neurological and psychiatric disorders such as chronic pain, Alzheimer's disease, major depression and schizophrenia. The ultracentrifuge will allow us to obtain fractions of subcellular organelles and to examine the interested proteins in each fraction, thus leading to insights into the underlying mechanisms. The requested equipment is an invaluable tool for the accomplishment of the research programs of the seven applicants and will have a significant impact on research progress at the Douglas Institute. Ultracentrifuge is an essential tool for any cutting-edge molecular neuroscience research environment and will strengthen the competitiveness of Douglas researchers nationally and internationally.