Research tools for studying local RNA translation at synapses

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Neurons are cells within the brain that transmit information to other nerve cells. Synapses are the contact points where one neuron communicates with another. My research program is aimed at understanding the mechanism by which neurons maintain synaptic connections. In particular, we want to understand how the production of proteins (i.e. the conversion of RNA to protein) by RNA-binding proteins (RBPs) occurs at synapses and how RBP activity is regulated. To study the production of proteins at the synapse and the regulation of RBP activity, fresh brain and spinal cord tissues are collected from mice. Harvested tissues are precisely cut to a 250 micron thickness with a vibratome machine, recovered in a special solution to keep them living, and then treated with different drugs or compounds. Using biochemical fractionation techniques, synapses are isolated from the fresh tissue slices and analyzed using a battery of biochemical and molecular approaches. To conduct these highly specialized experiments, we require the use of a vibratome machine that is dedicated for fresh tissues. The current vibratome that we are using is old and in disrepair. Currently, we are borrowing a new vibratome from another laboratory, but as my research team grows, the vibratome that we borrow is frequently not available. Additionally, there are logistical issues regarding where the vibratome is located. We must relocate our mice and supplies from our laboratory on the 3rd floor to the vibratome on the 5th floor to conduct our experiments. This is problematic because we must stop our time-sensitive workflow to brin our samples back to the 3rd floor for the biochemical isolation of synapses. As such, the logistics of coordinating our experiments have put our sample quality in jeopardy and slowed the progress of our projects. Thus, a vibratome is a critical and essential piece of equipment for my research program and to my trainees. This grant application is proposing the acquisition of a new vibratome for our studies. Our research program is taking an innovative and unique approach to understanding how synaptic connections are maintained and how synaptic connectivity impacts brain health and aging. Findings from our studies will act as a stepping-stone towards the long-term goal of our research program, which is to determine how protein synthesis maintains synaptic connections over the life-time of a neuron. The acquisition of a vibratome will allow us to push our research further so that we can have a better understanding of how synapses are maintained. We hope our findings will provide novel insights into the regulatory mechanisms that control local protein production and lead to the development of therapeutic approaches to treated age-related disorders, like amyotrophic lateral sclerosis (ALS), where synaptic loss is an early marker of these diseases.