Exploring synaptic remodeling with graphene optoelectronic probes

 描述（由申请人提供）：树突棘和突触的活性和可塑性是正常认知过程（如学习和记忆）的基础，也是大脑中复杂回路的基础。 树突棘是从树突轴发出的富含肌动蛋白的突起，包括大多数兴奋性突触的突触后末梢。 毫不奇怪，树突棘的异常与许多神经系统疾病有关，包括脆性X综合征、唐氏综合征、阿尔茨海默病、自闭症、精神分裂症和癫痫。 尽管棘和突触在中枢神经系统中的重要性，但调节这些结构的活性和可塑性的分子机制还没有很好地理解，这主要是因为目前缺乏在单个棘/突触水平上探测这些结构的可用技术。 此外，研究单个棘和突触的突触活动和可塑性的能力将为调节这些结构的功能和分子机制提供重要的见解。 我们正在开发具有集成石墨烯传感器和电极的新型神经元-神经胶质共培养微流体装置，并将其与扫描光电流显微镜结合，以亚突触分辨率检测和刺激脊柱可塑性（特定目标I）。 我们将使用 这项技术可以记录单个树突棘和突触的电特性，并检查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的活性、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（特异性目的II）。 我们将改变VASP的表达，并确定石墨烯探针对单个树突棘和突触电特性的影响。 此外，我们将确定这种蛋白质对突触可塑性的贡献。 提出的微流体平台的发展将是极大的兴趣和受益于神经生物学家提供了一个强大的技术，调查的机制，在一个单一的突触水平的电活动和可塑性的树突棘和突触。

 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.