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.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 该项目的目标是使用一种新的逆行追踪方法（我们称之为“逆行光动力学”）来确定神经节细胞类型的形态和生理学以及中心连接。我们使用罗丹明-葡聚糖逆行标记来自主要视网膜目标（上丘、前顶盖和 LGN）注射示踪剂的猕猴神经节细胞。正如逆行运输后所预期的那样，示踪剂被隔离在神经节细胞体和近端树突内的细胞器样结构中。这种颗粒标记本身并不能明确地靶向体外的特定细胞类型。然而，当在落射荧光照明下用显微镜观察标记细胞时，发光的细胞器似乎会破裂，在细胞质中产生烟花般的展示，并且释放的荧光团迅速扩散到整个树突树。同时，细胞质内荧光强度的大幅增加导致细胞内染料染色明亮且完整。光染色细胞在解剖学和生理学上保持活力；我们在体外针对形态上不同的类型进行细胞内记录和感受野特性分析。此外，通过使用生物素化形式的罗丹明葡聚糖，可以在组织固定后使用辣根过氧化物酶（HRP）组织化学永久恢复大量细胞的详细形态以进行解剖分析。这种方法使我们能够快速表征在 LGN 的主要视觉通路中投射的几个新神经节细胞群。其中一些细胞群表现出新颖的颜色对手特性，并将成为新研究项目的持续焦点。 其中一组，巨大的单层细胞，具有独特的光敏性，构成了实验室另一个项目的基础。我们还对视网膜中黑视蛋白反应细胞及其中央终端的进一步免疫组织化学研究感兴趣，并为此在丹麦招募了一位合作者。