Research tools for studying local RNA translation at synapses

神经元是大脑中向其他神经细胞传递信息的细胞。突触是一个神经元与另一个神经元进行交流的接触点。我的研究项目旨在了解神经元维持突触连接的机制。特别是，我们想了解RNA结合蛋白(RBPs)是如何在突触上产生蛋白质(即RNA到蛋白质的转换)的，以及RBP活性是如何受到调节的。为了研究突触蛋白的产生和RBP活性的调节，采集了小鼠新鲜的脑和脊髓组织。收获的组织用振动切割机精确切割成250微米厚，在特殊的溶液中恢复以保持它们的生命，然后用不同的药物或化合物进行治疗。使用生化分离技术，从新鲜的组织切片中分离突触，并使用一系列生化和分子方法进行分析。为了进行这些高度专业化的实验，我们需要使用专门用于新鲜组织的振动切割机。我们目前使用的振动器是旧的，年久失修。目前，我们正在从另一个实验室借来一个新的振荡器，但随着我的研究团队的壮大，我们借来的振动器经常是不可用的。此外，还有关于振动器位置的后勤问题。我们必须将我们的小鼠和用品从3楼的实验室搬到5楼的振动器进行实验。这是有问题的，因为我们必须停止时间敏感的工作流程，将我们的样本带回3楼进行突触的生化分离。因此，协调我们实验的后勤工作使我们的样品质量处于危险之中，并减缓了我们项目的进度。因此，对于我的研究项目和我的受训者来说，振动刀是一个关键和必不可少的设备。这项拨款申请建议购买一台新的振动器用于我们的研究。我们的研究项目正在采取一种创新和独特的方法来了解突触连接是如何维持的，以及突触连接如何影响大脑健康和衰老。我们的研究结果将作为迈向我们研究计划长期目标的垫脚石，该计划旨在确定蛋白质合成如何在神经元的生命周期中维持突触连接。获得振荡器将使我们能够进一步推动我们的研究，这样我们就可以更好地了解突触是如何维持的。我们希望我们的发现将为控制局部蛋白质生产的调控机制提供新的见解，并导致开发治疗与年龄相关的疾病的方法，如肌萎缩侧索硬化症(ALS)，突触丢失是这些疾病的早期标志。

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.