Ultracentrifugation for the biochemical characterization of biomarkers involved in neurobiological functions

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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.