Exploring synaptic remodeling with graphene optoelectronic probes

 描述(申请人提供)：树突棘和突触的活动和可塑性是正常认知过程(如学习和记忆)的基础，也是大脑中复杂电路的基础。树突棘是从树突干发出的富含肌动蛋白的突起，构成了兴奋性突触的大部分突触后终末。毫不奇怪，树突棘的异常与许多神经疾病有关，包括脆性X综合征、唐氏综合症、阿尔茨海默病、自闭症、精神分裂症和癫痫。尽管脊椎和突触在中枢神经系统中很重要，但调控这些结构活性和可塑性的分子机制在很大程度上是因为目前缺乏在单个脊椎/突触水平上探测这些结构的现有技术。此外，研究单个棘突和突触的突触活性和可塑性的能力将提供对调节这些结构的功能和分子机制的重要洞察。我们正在开发集成了石墨烯传感器和电极的新型神经元-胶质细胞共培养微流控设备，并将它们与扫描光电显微镜相结合，以检测和刺激突触下分辨率的脊髓可塑性(特定目标I)。我们将使用 这项技术记录了单个树突和突触的电特性，并检查了不同电刺激对这些结构的影响。由于肌动蛋白细胞骨架的重组被认为是树突棘和突触活性、可塑性和功能的基础，我们将探讨肌动蛋白结合蛋白Vasp在调节突触活性和可塑性中的作用(特定目标II)。我们将改变Vasp的表达，并用石墨烯探针确定对单个树突棘和突触的电学性质的影响。此外，我们还将确定该蛋白对突触可塑性的贡献。所提出的微流控平台的发展将是神经生物学家非常感兴趣和有益的，因为它提供了一种强大的技术来在单个突触水平上研究树突棘和突触的电活动和可塑性的机制。

 DESCRIPTION (provided by applicant): The activity and plasticity of dendritic spines and synapses underlie normal cognitive processes, such as learning and memory and are the basis for the complex circuitry found in the brain. Dendritic spines, which are actin-rich protrusions that emanate from the dendrite shaft, comprise most postsynaptic terminals of excitatory synapses. Not surprisingly, abnormalities in dendritic spines are associated with a number of neurological disorders, including Fragile-X syndrome, Down's syndrome, Alzheimer's disease, autism, schizophrenia, and epilepsy. Despite the importance of spines and synapses in the central nervous system, the molecular mechanisms that regulate the activity and plasticity of these structures are not well understood largely because of the current lack of available technologies for probing these structures at single spine/synapse levels. Furthermore, the capability to study synaptic activity and plasticity in individual spines and synapses would provide significant insight into the function and molecular mechanisms that regulate these structures. We are developing novel neuron-glia co-culture microfluidic devices with integrated graphene sensors and electrodes and combining them with scanning photocurrent microscopy to detect and stimulate spine plasticity at sub- synaptic resolution (Specific Aim I). We will use this technology to record electrical properties at individual dendritic spines and synapses and to examine the effects of different electrical stimuli on these structures. Since reorganization of te actin cytoskeleton is thought to underlie the activity, plasticity, and function of dendritic spine and synapses, we will explore the role of actin-binding protein VASP in regulating synaptic activity and plasticity (Specific Aim II). We will alter the expression of VASP and determine the effect on the electrical properties of individual dendritic spines and synapses with the graphene probes. Moreover, we will determine the contribution of this protein to synaptic plasticity. The development of the proposed microfluidic platforms will be of great interest and benefit to neurobiologists by providing a powerful technology for investigating the mechanisms that underlie the electrical activity and plasticity of dendritic spines and synapses at a single synaps level.