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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 该项目的目标是使用我们称为“逆行光动力学”的新逆行追踪方法来确定神经节细胞类型的形态学和生理学以及中枢连接。我们已经使用罗丹明-葡聚糖逆行标记猕猴神经节细胞的示踪剂注射在主要的视网膜目标：上级丘，pretectum，和LGN。正如逆行转运后预期的那样，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。这种颗粒标记单独不允许明确靶向特定的细胞类型在体外。然而，当标记的细胞在显微镜下观察落射光照明，发光的细胞器似乎爆裂在细胞质中形成一个烟花状的展示释放的荧光团迅速扩散到整个树枝状树。同时，细胞质内荧光强度的大幅增加产生明亮且完整的细胞内染料染色。光染色的细胞在解剖学和生理学上保持活力;我们针对体外形态学上不同的类型进行细胞内记录和感受野特性分析。此外，通过采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）组织化学来永久恢复大量细胞的详细形态以用于解剖分析。这种方法使我们能够快速表征几个新的神经节细胞群，项目在主要的视觉通路的LGN。其中一些细胞群显示出新颖的颜色对手属性，并将成为新研究项目的持续重点。 其中一组，巨大的单层细胞，具有独特的感光性，并形成了实验室中另一个项目的基础。我们也有兴趣在进一步的免疫组织化学研究的黑视素反应细胞在视网膜以及他们的中央终端，并已招募了合作者在丹麦为此。