Functional Dissection of New Retinal Circuits

新视网膜回路的功能解剖

• 批准号: 10368098
• 负责人: Z JIMMY ZHOU
• 金额: $ 60.46万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10368098
• 关键词: Address; Amacrine Cells; Cells; Classification; Cone; Data; Disease; Dissection; Electrophysiology (science); Eye diseases; Feedback; Genes; Genetic; Glutamates; Goals; Health; Human; Image; Inner Nuclear Layer; Light; Link; Modeling; Mus; Neurons; Optics; Output; Physiology; Preparation; Property; Public Health; Research; Retina; Role; SLC17A8 gene; Shapes; Synapses; Synaptic Transmission; Techniques; Testing; Th2 Cells; Transfection; Viral; base; cell type; ganglion cell; in vivo; innovation; insight; neural circuit; neuronal circuitry; novel; optogenetics; patch clamp; receptive field; relating to nervous system; response; synaptic function; tool; two-photon

The goal of this application is to understand the connectivity and function of a unique dual-transmitter amacrine cell circuit, in an effort to gain a comprehensive understanding of the functional organization of local neuronal circuits in the inner mammalian retina. Our recent studies discovered a number of intriguing synaptic and dendritic properties of the vGluT3 amacrine cell (GAC) network (Lee et al., Neuron 84:1049-64, 2014; Lee et al. Neuron 90: 27-34, 2016; Chen et al., PNAS 114:11518-11523, 2017), which offer an exciting opportunity to use this new model circuit to address many fundamental, long-standing questions regarding amacrine cell circuitry and function. Here, we propose to take a systematic approach to uncovering the connectivity and function of the entire local circuit of GACs, from the inputs from upstream bipolar cell types to the outputs to distinct downstream ganglion cell and amacrine cell types. The proposed study will focus on identifying new synaptic circuits between GACs and their synaptic partners and uncovering novel functional roles of GACs in shaping the receptive field properties of their downstream targets. Using the GAC circuit as a central link, we also aim to understand how the GAC circuit connectivity may help delineate the organization of other local circuits in the retina. There are three specific aims in this proposal: (1) to understand the synaptic connectivity and synaptic interactions between cone bipolar cell (CBC) types and GACs, (2) to understand function of a new glycinergic output circuit from GACs to ganglion cells, (3) to understand network interactions between GACs and other amacrine cell types. We will take an innovative approach to this goal by combining state-of-art electrophysiological (dual patch clamp in the inner nuclear layer of the wholemount mouse retina), optical (simultaneous two-photon imaging of local dendritic responses and patch clamp), genetic (in vivo viral transfection of genes in selected single neurons), and optogenetic techniques in a robust whole-mount preparation of vGluT3-Cre mouse retina. These techniques have been well tested in our recent studies and have helped us generate a substantial amount of preliminary data that are critical for the proposed study. The combination of these powerful tools with a unique, well-defined preparation offers an opportunity rarely attainable in other parts of the CNS. Results from this proposed research will shed important light on retinal circuit organization and function in health and disease.

此应用程序的目标是了解独特的双发射器的连接性和功能 无长突细胞回路，以努力全面了解局部的功能组织， 哺乳动物视网膜内层的神经元回路。我们最近的研究发现了一些有趣的突触 和vGluT 3无长突细胞（GAC）网络的树突特性（Lee等，Neuron 84：1049-64，2014; Lee 等人Neuron 90：27-34，2016; Chen等人，PNAS 114：11518-11523，2017），其提供了令人兴奋的机会 使用这种新的模型电路来解决许多关于无长突细胞的基本的，长期存在的问题， 电路和功能。在这里，我们建议采取一种系统的方法来揭示连通性， 从上游双极单元类型的输入到输出， 不同的下游神经节细胞和无长突细胞类型。拟议的研究将侧重于确定新的 GACs和它们的突触伙伴之间的突触回路，并揭示了GACs在 形成其下游目标的感受野特性。使用GAC电路作为中心链路，我们 我还希望了解GAC电路连接如何帮助描述其他本地电路的组织 在视网膜上。本研究有三个具体目标：（1）了解突触连接， 突触之间的相互作用锥双极细胞（CBC）类型和GACs，（2）了解功能的新的 （3）了解从GACs到神经节细胞的甘氨酸能输出回路，（4）了解GACs之间的网络相互作用， 其他无长突细胞类型。我们将采取创新的方法来实现这一目标， 电生理学（在整个小鼠视网膜的内核层中的双膜片钳）、光学 （局部树突状反应和膜片钳的同时双光子成像），遗传（体内病毒 在选定的单个神经元中的基因转染），以及在稳健的整体安装中的光遗传学技术。 vGluT 3-Cre小鼠视网膜的制备。这些技术在我们最近的研究中得到了很好的测试， 帮助我们产生了大量的初步数据，这些数据对拟议的研究至关重要。的 将这些强大的工具与独特的、定义明确的准备相结合，提供了一个难得的机会 在中枢神经系统的其他部分。这项研究的结果将为视网膜回路提供重要的线索 健康和疾病的组织和功能。