Optical Determination of Neuronal Signaling and Circuits

描述(由申请人提供)：本提案寻求支持从HHMI Janelia农场研究园区聘请一名年轻的丈夫和IFE团队，他们将为Vollum研究所增加巨大的力量。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电路和信号传递的问题，这是Vollum现有优势的补充。钟博士最近完成了与卡雷尔·斯沃博达和埃里克·贝齐格的博士后研究，贝齐格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完成了博士后研究。我们的目的是利用这些PSO资金来帮助抵消他们复杂的微观需求的成本，以便可用的启动资金可以用于较长期的承诺。这两名调查人员将与现有的Vollum教职员工进行广泛的交叉，人们对聘用他们的前景普遍感到兴奋。PSO资金对于确保他们成功过渡到独立至关重要。传统的电生理和生化方法虽然在解决许多问题方面很强大，但缺乏必要的时空分辨率来解决关键问题。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。他将开发将Palm与电子显微镜相结合的方法来分析PKA在突触脊椎中的定位规律，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。MAO还使用先进的成像技术，结合分子和生理学工具，如激光扫描光刺激(LSPs)、通道视紫红质辅助电路映射(CRACM)和遗传编码的钙传感器，来绘制基底节奖励相关行为的局部电路。她最初将专注于比较大脑皮质和丘脑不同区域的输入如何不同地影响不同隔室内和不同隔室之间的纹状体神经元活动。一个更长期的目标将是将行为不同阶段的回路变化(例如，目标导向型与习惯性型)描述为成瘾的动物模型。这套招募将允许Vollum进一步在突触调制和信号传递方面的卓越地位，这是其计划的中心焦点，并扩展到神经元电路领域。 与公共健康相关：突触功能和可塑性由复杂的信号转导网络控制。尽管已经确定了这些网络的许多组成部分，但仍然面临着理解它们如何对突触信号机制做出贡献的挑战。解析真实的突触信号事件需要比以前更高的空间和时间分辨率。钟博士和毛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.