Development of retinal ganglion cell types

视网膜神经节细胞类型的发育

• 批准号: 8373861
• 负责人: DAVID A FELDHEIM
• 金额: $ 42万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8373861
• 关键词: Ablation; Address; Adult; Affect; Amaze; Axon; Biological Assay; Brain; Cell Differentiation process; Cell Proliferation; Cell physiology; Cells; Cellular Morphology; Code; Color; Commit; 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; 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; base; 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

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. PUBLIC HEALTH RELEVANCE:! In this grant we propose experiments designed to understand the mechanisms by which a common pool of retinal ganglion cell progenitors gives rise to distinct functional types of mature RGCs, each of which participates in a unique neural circuit that encodes a specific aspect of the visual scene: experiments proposed in this grant use a combination of mouse molecular genetics and state of the art neural recording technology to determine the role of the transcription factors Isl2 and Satb2 in RGC functional specification. Upon completion of the proposed aims, we have will insights into how different retinal circuits arise during development: increased knowledge of the molecular control of RGC development is likely to have practical benefits in understanding the consequences of diseases that affect RGC function. Successful therapies for 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）：该提案的目标是了解用于创建哺乳动物视网膜电路的机制。具体来说，实验旨在了解转录因子 Islet-2 (Isl2) 和特定的富含 AT 的 DNA 结合蛋白 2 (Satb2) 在不同类型视网膜神经节细胞规范中的作用 （RGC）。 RGC 是视网膜的输出神经元，会受到青光眼和视神经发育不全等多种视网膜疾病的影响。哺乳动物视网膜中有多种功能类型的 RGC，每种类型都参与编码视觉场景的特定方面（例如运动、空间图案或颜色）的视网膜回路。这种功能特异性源自不同的 RGC 形态以及与其他细胞类型的选择性突触形成；然而，两者在开发过程中如何建立仍不清楚。我们的实验结合了小鼠转基因技术、体内生理电记录和成像技术以及解剖追踪。我们关注 Isl2 和 Satb2，因为它们的表达仅限于特定的 RGC 类别，从而表明特定的功能。 在目标 1 中，我们将使用形态学和生理学标准确定哪些类型的 RGC 表达 Isl2 和 Satb2。我们的假设是每个 TF 指定 RGC 类型的共同感受野属性；例如，Isl2+ RGC 是开启和关闭的 RGC，而 Satb2 RGC 可能构成方向选择性细胞类型。目标 1a 是确定在定义的 RGC 子集中表达 GFP 的小鼠中 Isl2 的表达。目标 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。 在目标 2 中，我们将通过从视网膜特异性去除 Isl2 或 Satb2 并在发育中的 RGC 中异位表达它们来确定 Isl2 和 Satb2 对于 RGC 分化为其正常细胞类型是否是必要和充分的。然后，我们将使用各种生理和形态学标准确定所得 RGC 的形态学和生理学特性。 我们研究 RGC 分化的方法将利用我们对不同基因标记的 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。 这将进一步加深我们对不同视网膜回路在发育过程中如何产生的理解；这反过来可能对了解影响 RGC 功能的疾病的后果有实际的好处。此外，治疗视网膜疾病的成功疗法将取决于 受损视觉回路的恢复，使了解这些回路的发展对于设计治疗策略和评估功能恢复至关重要。 公共卫生相关性：！在这笔资助中，我们提出了一些实验，旨在了解视网膜神经节细胞祖细胞的共同池产生不同功能类型的成熟 RGC 的机制，每个 RGC 都参与编码视觉场景特定方面的独特神经回路：本次资助中提出的实验结合了小鼠分子遗传学和最先进的神经记录技术来确定转录因子 Isl2 和 Satb2 在 RGC 功能规范中的作用。完成所提出的目标后，我们将深入了解不同的视网膜回路在发育过程中是如何产生的：增加对 RGC 发育的分子控制的了解可能对理解影响 RGC 功能的疾病的后果有实际的好处。视网膜疾病的成功治疗将取决于受损视觉回路的恢复，因此了解这些回路的发展对于设计治疗策略和评估功能恢复至关重要。