Development of retinal ganglion cell types

视网膜神经节细胞类型的发育

基本信息

批准号：
8879150
负责人：
DAVID A FELDHEIM
金额：
$ 37.36万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA CRUZ
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8879150
关键词：
Ablation Address Adult Affect Amaze Axon Biological Assay Brain Cell Differentiation process Cell Proliferation Cell physiology Cells Cellular Morphology Code Color DNA-Binding Proteins Development Disease Ectopic Expression Electrodes Electroporation Embryo Exhibits Future Genes Glaucoma Goals Grant Imaging Techniques In Vitro Individual Knockout Mice Knowledge Label Lead Left Life Measures Mitosis Mitotic Molecular Molecular Genetics Morphology Motion Mus Neurons Normal Cell Optic Nerve Outcomes Research Output Pattern Physical shape Physiological Physiology Proliferation Marker Property Proteins Protocols documentation Recovery of Function Replacement Therapy Retina Retinal Retinal Diseases Retinal Ganglion Cells Role Specific qualifier value Specificity Spinal Cord Staging Structure Techniques Technology Testing Therapeutic Transgenic Mice Viral Visual anatomical tracing base brain circuitry cell type design embryonic stem cell in vivo insight islet mutant nerve stem cell neural circuit novel strategies prevent progenitor receptive field relating to nervous system research study response restoration stem synaptogenesis transcription factor visual information

项目摘要

DESCRIPTION (provided by applicant): The goal of this proposal is to understand the mechanisms used to create the circuitry of the mammalian retina. Specifically, experiments are aimed toward understanding the role of the transcription factors Islet-2 (Isl2) and specific AT-rich DNA-binding protein 2 (Satb2) in the specification of distinct types of retinal ganglion cells (RGCs). RGCs are the output neurons of the retina and are affected in several retinal diseases such as glaucoma and optic nerve hypoplasias. There are numerous functional types of RGCs in the mammalian retina, each participating in a retinal circuit that encodes a specific aspect of the visual scene, such as motion, spatial patterns, or color. This functional specificity is derive from distinct RGC morphology and selective synapse formation with other cell types; however, how both are established during development remains unclear. Our experiments use a combination of mouse transgenic technology, in vivo physiological electrical recording and imaging techniques, and anatomical tracing. We focus on Isl2 and Satb2 because their expression is restricted to specific RGC classes, thereby suggesting a specific function. In Aim 1 we will determine which types of RGCs express Isl2 and Satb2 using both morphological and physiological criteria. Our hypothesis is that each TF specifies a common receptive field property of RGC types; for example, that Isl2+ RGCs are On and Off RGCs, while Satb2 RGCs may constitute direction selective cell types. Aim 1a is to determine Isl2 expression in mice expressing GFP in defined RGC subsets. Aims 1b and c together aim to classify all Isl2+ RGCs based on morphological and physiological criteria. In Aim 2 we will determine if Isl2 and Satb2 are necessary and sufficient for the RGCs to differentiate into their normal cell types by specifically removing Isl2 or Satb2 from the retina and by ectopically expressing them in developing RGCs. We will then determine the morphological and physiological properties of the resulting RGCs using a variety of physiological and morphological criteria. Our approach to study RGC differentiation will use our knowledge of the structure and function of different genetically-labeled RGC types to identify transcription factors that lead to their differentiation. This will further our understanding of how different retinal circuits arise during development; thi in turn is likely to have practical benefits in understanding the consequences of diseases that affect RGC function. In addition, successful therapies for treating retinal diseases will depend on the restoration of damaged visual circuits, making knowledge of the development of these circuits essential for designing therapeutic strategies and assessing functional recovery.

描述（由申请人提供）：该提案的目的是了解用于创建哺乳动物视网膜电路的机制。具体而言，实验旨在了解转录因子ISLET-2（ISL2）和特定富含DNA结合蛋白2（SATB2）在不同类型的视网膜神经节细胞中的作用（RGC）。 RGC是视网膜的输出神经元，在几种视网膜疾病（例如青光眼和视神经下腹膜）中受到影响。哺乳动物视网膜中有许多功能类型的RGC，每种RGC都参与了编码视觉场景特定方面的视网膜电路，例如运动，空间模式或颜色。该功能特异性来自不同的RGC形态和其他细胞类型的选择性突触形成。但是，在开发过程中如何建立两者尚不清楚。我们的实验结合了小鼠转基因技术，体内生理电气记录和成像技术以及解剖过程。我们专注于ISL2和SATB2，因为它们的表达仅限于特定的RGC类，从而暗示了特定功能。在AIM 1中，我们将使用形态和生理标准确定哪种类型的RGC表达ISL2和SATB2。我们的假设是每个TF指定RGC类型的常见野外特性。例如，ISL2+ RGC在RGC中打开和关闭，而SATB2 RGC可能构成方向选择性细胞类型。 AIM 1A是确定在定义的RGC子集中表达GFP的小鼠中的ISL2表达。 AIMS 1B和C共同旨在根据形态和生理标准对所有ISL2+ RGC进行分类。在AIM 2中，我们将确定ISL2和SATB2是否需要且足以使RGC通过专门将ISL2或SATB2从视网膜中删除，并在开发RGC中表达它们来区分其正常细胞类型。然后，我们将使用各种生理和形态标准来确定所得RGC的形态和生理特性。我们研究RGC分化的方法将利用我们对不同遗传标记的RGC类型的结构和功能的了解来识别导致其分化的转录因子。这将进一步了解我们在开发过程中如何出现不同的视网膜电路。反过来，这可能在理解影响RGC功能的疾病的后果方面具有实际好处。此外，成功治疗视网膜疾病的疗法将取决于恢复受损的视觉电路，了解这些电路的发展，对于设计治疗策略和评估功能恢复至关重要。