Optical Determination of Neuronal Signaling and Circuits

描述（由申请人提供）：本提案寻求支持雇用一个年轻的丈夫和ife团队从HHMI珍妮利亚农场研究校园谁将增加巨大的力量，伏勒姆研究所。钟海宁博士和毛天一博士利用新开发的光学方法来解决涉及神经回路和信号的问题，这是对Vollum现有优势的补充。钟博士最近完成了与Karel Svoboda和Eric Betzig的博士后研究，他们是光活化定位显微镜（PALM）的共同发明者。毛博士还在斯沃博达实验室做博士后研究。我们的目的是利用这些PSO资金来帮助抵消其复杂的微观需求的成本，以便可用的启动资金可以用于更长期的承诺。这两名调查人员将与现有的沃勒姆教师有广泛的交集，人们对雇用他们的前景普遍感到兴奋。支持和平行动的资金对于确保其成功过渡到独立至关重要。传统的电生理和生物化学方法，虽然在解决许多问题的强大，缺乏必要的时空分辨率解决关键问题。钟博士利用新开发的成像方法，超越了传统光学显微镜的衍射极限，探索突触的信号通路。他将联合收割机与电子显微镜相结合，研究PKA在突触棘中定位的调控，并使用双光子荧光寿命成像（FLIM）来测量PKA与相关蛋白的直接相互作用。毛博士还使用先进的成像技术，以及分子和生理学工具，如激光扫描光刺激（LSPS），通道视紫红质辅助电路映射（CRACM），以及基因编码的钙传感器，以绘制基底神经节中与奖励相关的行为的局部回路。她最初将专注于比较来自皮层和丘脑不同区域的输入如何差异地影响不同隔室内部和之间的纹状体神经元活动。长期目标将是通过行为的不同阶段来表征电路变化（例如，目标导向与习惯性）作为成瘾的动物模型。这一系列的招募将使伏伦在突触调制和信号传导方面进一步发挥其卓越的作用，这是其程序的中心焦点，并延伸到神经元回路领域。 公共卫生相关性：突触功能和可塑性由复杂的信号传导网络控制。虽然这些网络的许多组成部分已经被确定，但挑战仍然是了解它们如何有助于突触信号机制。解析真实的突触信号事件需要比先前可用的更大的空间和时间分辨率。Zhong和Mao博士利用先进的成像方法来解决突触调节中的关键问题。

DESCRIPTION (provided by applicant): This proposal seeks support for hiring a young husband and ife team from the HHMI Janelia Farm Research Campus who will add great strength to the Vollum Institute. Drs. Haining Zhong and Tianyi Mao utilize newly developed optical approaches to solve problems involving neural circuits and signaling, an area complementary to the Vollum's existing strengths. Dr. Zhong recently completed his postdoctoral fellowship with Karel Svoboda and Eric Betzig, co-inventor of photoactivated localization microscopy (PALM). Dr. Mao also did her postdoctoral fellowship in the Svoboda lab. Our intent is to use these PSO funds to help offset the costs of their sophisticated microscopic needs so that the available start up funds can be used for longer term commitment. These two investigators will have extensive intersections with existing Vollum faculty, and there is widespread excitement at the prospect of hiring them. PSO funds are critical for insuring their successful transition to independence. Traditional electrophysiological and biochemical approaches, although powerful in addressing many questions, lack the necessary spatiotemporai resolution for addressing key questions. Dr. Zhong utilizes newly developed imaging methodologies that extend beyond the diffraction limit of conventional light microscopy to explore signaling pathways at the synapse. He will develop methods to combine PALM with electron microscopy to analyze the regulation of PKA localization in synaptic spines and use two-photon fluorescence lifetime imaging (FLIM) to measure direct interaction of PKA with associated proteins.Dr. Mao also uses advanced imaging techniques, together with molecular and physiology tools such as laser scanning photostimulation (LSPS), channelrhodopsin-assisted circuitry mapping (CRACM), and genetically encoded calcium sensors to map local circuits underlying reward-related behaviors in the basal ganglia. She initially will focus on comparing how the inputs from various areas of the cortex and the thalamus differentially influence striatal neuronal activities within and between different compartments. A longer-term goal will be to characterize circuit changes through different stages of behavior {e.g., goal-directed vs. habitual) as animal models of addiction. This set of recruitments would allow the Vollum to further its preeminence in synaptic modulation and signaling, the central focus of its program, and extend into the area of neuronal circuitry. PUBLIC HEALTH RELEVANCE: Synaptic function and plasticity are controlled by complex signal ransduction networks. Although many components of these networks have been identified, the challenge remains to understand how they contribute to synaptic signaling mechanisms. Resolving the authentic synaptic signaling events requires greater spatial and temporal resolution than previously available. Drs. Zhong and Mao utilize advanced imaging approaches to address key questions in synaptic regulation.