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.

该应用程序的目的是了解唯一双发送器的连接性和功能 无摩碱细胞电路，以便全面了解本地的功能组织 内部哺乳动物视网膜中的神经元电路。我们最近的研究发现了许多有趣的突触 Vglut3 amacrine细胞（GAC）网络（Lee等人，Neuron 84：1049-64，2014; Lee 等。 Neuron 90：27-34，2016; Chen等人，PNAS 114：11518-11523，2017），提供了令人兴奋的机会 要使用这个新的模型电路来解决有关无氨基氨基网细胞的许多基本，长期存在的问题 电路和功能。在这里，我们建议采用一种系统的方法来揭示连接性和 GAC的整个本地电路的功能，从上游双极单元类型到输出的输入到输出 独特的下游神经节细胞和无长熟细胞类型。拟议的研究将着重于确定新的 GACS与其突触伙伴之间的突触电路，并发现GAC的新功能作用 塑造其下游目标的接受场特性。使用GAC电路作为中心链接，我们 还旨在了解GAC电路连接如何帮助描述其他本地电路的组织 在视网膜中。该提案中有三​​个具体目标：（1）了解突触连通性和 锥双极细胞（CBC）类型与GAC之间的突触相互作用，（2）了解新的功能 （3）了解GAC和GAC之间的网络相互作用 其他无链氨酸细胞类型。我们将通过结合最先进来对此目标采取创新的方法 电生理学（电网的内部核层中的双贴片夹），光学 （同时对局部树突反应和斑块夹的两光子成像），遗传（体内病毒 在选定的单个神经元中转染基因），并在强大的整体上转染光遗传学技术 VGLUT3-CRE小鼠视网膜的制备。这些技术在我们最近的研究中都经过了很好的测试， 已经帮助我们产生了大量的初步数据，这对于拟议的研究至关重要。这 这些功能强大的工具与独特，定义明确的准备工作的结合提供了很少能实现的机会 在中枢神经系统的其他部分。这项拟议的研究的结果将在视网膜电路上散发出重要的灯光 健康与疾病的组织和功能。