ANATOMY AND PHYSIOLOGY OF NOVEL GANGLION CELL TYPES IN MACAQUE RETINA

猕猴视网膜中新型神经节细胞的解剖学和生理学

• 批准号: 8357583
• 负责人: DENNIS MICHAEL DACEY
• 金额: $ 15.66万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357583
• 关键词: Anatomy; Cell Count; Cells; Color; Cytoplasm; Dendrites; Denmark; Diffuse; Dyes; Fluorescence; Funding; Goals; Grant; Histocytochemistry; Horseradish Peroxidase; In Vitro; Injection of therapeutic agent; Label; Lighting; Macaca; Methods; Monkeys; Morphology; National Center for Research Resources; Organelles; Particulate; Physiology; Population; Primates; Principal Investigator; Property; Research; Research Infrastructure; Research Project Grants; Resources; Retina; Retinal; Source; Staining method; Stains; Structure; Time; Tissue Fixation; Tracer; Trees; United States National Institutes of Health; Visual Pathways; base; cell type; cost; fluorophore; ganglion cell; interest; melanopsin; neuronal cell body; novel; receptive field; retrograde transport; rhodamine dextran; superior colliculus Corpora quadrigemina

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The goal of this project is to determine the morphology and physiology and central connections of ganglion cell types using a new retrograde tracing method that we have called 'retrograde photodynamics'. We have used rhodamine-dextrans to retrogradely label macaque monkey ganglion cells from tracer injections in the major retinal targets: the superior colliculus, pretectum, and LGN. As expected after retrograde transport, the tracer is sequestered in organelle-like structures within ganglion cell bodies and proximal dendrites. This particulate labeling alone does not allow unambiguous targeting of specific cell types in vitro. However when labeled cells were observed microscopically under epifluorescent illumination, the glowing organelles seem to burstcreating a fireworks-like display in the cytoplasmand the liberated fluorophore rapidly diffuses throughout the dendritic tree. At the same time, a large increase in fluorescence intensity within the cytoplasm gives rise to a bright and complete intracellular dye stain. Photostained cells remain anatomically and physiologically viable; we target morphologically distinct types in vitro for intracellular recording and analysis of receptive field properties. Further, by employing the biotinylated form of rhodamine dextran, it is possible to use horseradish peroxidase (HRP) histochemistry after tissue fixation to permanently recover the detailed morphology of large numbers of cells for anatomical analysis. This method enables us to rapidly characterize several new ganglion cell populations that project in the primary visual pathway to the LGN. Some of these cell groups show novel color-opponent properties and will be a continuing focus of new research projects. One of these groups, the giant monostratified cells, are uniquely photosensitive and form the basis for another project in the lab. We are also interested in further immunohistochemical studies of melanopsin-reactive cells in the retina as well as their central terminals, and have enlisted a collaborator in Denmark to this end.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 该项目的目的是使用我们称为“逆行光动力学”的新逆行跟踪方法来确定神经节细胞类型的形态和生理和中心连接。我们已经使用若丹明 - 滴力抗激逆，从主要视网膜靶标的示踪剂注射中逆行标记了猕猴的猴子神经节细胞：上胶胶，预性和LGN。正如逆行转运后所预期的那样，示踪剂在神经节细胞体内的细胞器样结构和近端树突中被隔离。仅此颗粒物标记不允许在体外明确靶向特定的细胞类型。然而，当在落入散荧光照明下观察到标记的细胞时，发光的细胞器似乎在细胞质中形成了烟花样的表现，而释放的荧光团在整个树突状树中迅速扩散。同时，细胞质内的荧光强度大大增加会导致明亮而完整的细胞内染料染色。光固化的细胞在解剖学和生理上保持生存。我们靶向体外形态上不同类型的类型，用于对接受场特性的细胞内记录和分析。此外，通过采用若丹明葡萄糖的生物素化形式，可以在组织固定后使用辣根过氧化物酶（HRP）组织化学来永久恢复大量细胞的详细形态进行解剖分析。这种方法使我们能够迅速表征在LGN主要视觉途径中投射的几个新的神经节细胞种群。其中一些细胞组显示出新颖的颜色 - 重点特性，并将成为新研究项目的持续重点。 这些组之一，即巨型单层化细胞，具有独特的光敏性，并构成了实验室中另一个项目的基础。我们还对视网膜及其中央末端的黑素反应性细胞的进一步免疫组织化学研究感兴趣，并已将合作者纳入丹麦。