Regulation of Neural Circuitry by Cypin

对于正常的神经元功能，神经元必须有正确数量的输入树突，这些树突看起来像树上的树枝。目前关于这些分支的模式是如何确定的，或者当一个人学习时这些分支是如何变化的，我们知之甚少。为了形成新的树突分支，必须产生一个细胞骨架，就像手指上的骨架一样。费尔斯坦博士发现了一种名为cypin的蛋白质，它在形成这种细胞骨架以及树突的过程中起着关键作用。据推测，如果没有cypin，神经元将无法正常形成，大脑将无法正常运行。利用培养皿中的神经细胞，本科生和研究生将进行实验，以了解cypin产生功能神经元的机制。通过研究cypin如何赋予神经细胞形状，以及这些神经细胞如何整合成简单的电路，这项研究将有助于我们理解在学习等事件中电路可能被修改的原理。费尔斯坦博士和他的同事将包括一批不同的本科生和研究生，并将与波多黎各大学建立一个交流项目。这项建议还包括通过制作视频系列《教我神经科学》和培训K-12教师来激发年轻一代(K-12)学生对神经科学的兴趣。费尔斯坦博士希望建立一个项目，通过研讨会、研讨会和交流项目将神经科学带到波多黎各的社区。目前工作的具体目标是评估cypin及其结合伙伴PSD-95如何影响神经回路动力学。实验旨在通过在细胞培养中通过病毒介导的基因表达改变cypin水平来确定cypin降低突触PSD-95的机制。将评估蛋白酶体在Cypin介导的PSD-95变化中的作用。此外，还将确定Cypin和PSD-95水平是否会影响体外神经回路的功能和活性。费尔斯坦博士拥有一套全面的工具，可以在分子水平上操纵cypin的功能，并将评估cypin水平变化对树突数量、脊椎数量和大小的影响。实验将利用电生理学技术来确定cypin是否会影响神经元信号。这项拟议的工作使用跨学科的方法--分子/细胞、生化和电生理学--来了解神经元的形态变化如何导致突触功能的变化，比以前的细胞培养工作有了很大的进步，并定义了一种机制，通过这种机制，cypin调节树突发生并确定对神经电路的影响。

For proper neuron function, the neuron must have the correct number of input dendrites, which look like branches on a tree. Very little is currently known about how the pattern of these branches is determined or how these branches change when a person learns. To make new dendrite branches, a cytoskeleton must be produced, much like the skeleton in fingers. Dr. Firestein discovered a protein called cypin that plays a critical role in making this cytoskeleton, and hence dendrites. It is hypothesized that without cypin, neurons will not form properly, and the brain will not function correctly. Using nerve cells in a dish, undergraduate and graduate students will perform experiments to understand the mechanism by which cypin acts to yield functioning neurons. By investigating how cypin gives nerve cells their shape and how these nerve cells integrate into simple circuits, this research will aid in our understanding of principles by which circuits may be modified during events, such as learning. Dr. Firestein and colleagues will include a diverse group of undergraduate and graduate students and will establish an exchange program with University of Puerto Rico. This proposal also encompasses activities to excite a younger generation of students (K-12) about neuroscience by producing a video series "Teach Me Neuroscience" and training K-12 teachers at the bench. It is Dr. Firestein's hope to establish a program to bring neuroscience to the community in Puerto Rico via seminars, workshops, and exchange programs. The specific goal of the current work is to evaluate how cypin and its binding partner PSD-95 affect neural circuit dynamics. Experiments aim to determine the mechanism by which cypin decreases synaptic PSD-95 using viral-mediated gene expression in cell culture to alter cypin levels. The role of the proteasome in cypin-mediated changes in PSD-95 will be assessed. It will also be determined whether cypin and PSD-95 levels affect function and activity of neural circuits in vitro. Dr. Firestein has a comprehensive set of tools available to manipulate cypin functionality at the molecular level and will assess effects of altered cypin levels on dendrite number, spine number, and size. Experiments will make use of electrophysiology techniques to determine whether cypin affects neuronal signaling. The proposed work uses interdisciplinary approaches - molecular/cellular, biochemical, and electrophysiological - to understand how morphological changes to neurons result in changes in synaptic function, making a large advance from previous cell culture work and defining a mechanism by which cypin acts to regulate dendritogenesis and determine effects on neural circuitry.