Early retinal development in pigmented and albino mice: Factors in the CMZ and RPE controlling RGC specification

色素沉着和白化小鼠的早期视网膜发育：CMZ 和 RPE 控制 RGC 规范的因素

• 批准号: 10163186
• 负责人: Carol A. Mason
• 金额: $ 53万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10163186
• 关键词: Albinism; Animals; Automobile Driving; Axon; Binocular Vision; Biological Models; Brain; CCND2 gene; Cell Cycle; Cell Cycle Regulation; Cell Lineage; Cell division; Cells; Communication; Contralateral; Dependence; Development; Distal; Embryo; Ensure; Event; Eye; Eye Development; Fiber; Generations; Genes; Genetic; Genetic Diseases; Genetic Transcription; Goals; Grant; Human; Hypopigmentation; Image; Ipsilateral; Lead; Ligands; Link; Location; Melanins; Melanogenesis; Modeling; Molecular; Mus; Neural Retina; Neurons; Optic Chiasm; Optic Nerve; Pathway interactions; Pattern; Peripheral; Phenotype; Pigments; Process; Production; Property; Regulation; Regulator Genes; Replacement Therapy; Reporter; Retina; Retinal Ganglion Cells; Retinal Pigments; Role; Route; Side; Signal Transduction; Source; Specific qualifier value; Structure of retinal pigment epithelium; Study models; System; Technology; Time; Vision; Vision Disorders; Visual Pathways; Visual system structure; WNT Signaling Pathway; Work; albino mouse; axon regeneration; base; cell fate specification; cell replacement therapy; comparative; genetic manipulation; injured; insight; migration; monolayer; neurogenesis; neuronal circuitry; novel; programs; public health relevance; receptor; segregation; single-cell RNA sequencing; spatiotemporal; stem cells; transcription factor; transcriptomics

PROJECT SUMMARY Retinal ganglion cells (RGCs) from each eye extend their axons ipsi- and contralaterally to the brain to establish the circuit for binocular vision. The lack of appropriate sidedness of RGC projections is debilitating in genetic disorders such as albinism, in which hypopigmentation of the retinal pigment epithelium (RPE) is linked to optic nerve misrouting and therefore altered stereo vision. Progress on this grant includes the identification of transcriptional regulators of the specification and differentiation of ipsi and contra RGCs; demonstration of the ventral ciliary margin zone (CMZ) as a source of ipsi RGCs; expression of the cell cycle regulator Cyclin D2 in the ventral CMZ; and dependence of the ipsi RGC projection on Cyclin D2. In the albino retina, these processes and the cellular integrity of the RPE, are disrupted, making the albino an excellent comparative model for the proposed studies. Here we explore a novel role for the CMZ in neural retinal development and RGC specification, and seek to uncover potential signaling networks for establishing proper RGC connections. We propose to combine studies of neurogenesis and fate mapping with transcriptomics of the CMZ, RPE, and neural retina to gain mechanistic insight into how loss of melanin in the RPE of albino animals causes a shift in cell fate from ipsi to contra during the establishment of the binocular circuit. To this end, we will establish a role for Cyclin D2 in the pace of the cell cycle, plane of cell division, and migration from the CMZ to the neural retina (Aim 1); study signaling between the RPE and CMZ, starting with the Wnt pathway, as a route to regulate RGC fate (Aim 2); identify the transcriptional networks that regulate RGC cell fate by performing single-cell RNA-Seq of the CMZ, neural retina, and RPE (Aim 3). Throughout, we will compare albino and pigmented retina. We hypothesize that events in the CMZ control timing of neurogenesis, which specifies RGC projection fate and that factors in the RPE provide directives to the CMZ and neural retina for ipsi/contra RGC fate acquisition. Significance: Probing early neural retinal development and parsing regulators of neurogenesis and cell fate emanating from the CMZ and RPE offer a novel mechanistic entry point to the long-standing enigma of how pigment and the RPE influences cell fate and RGC axon segregation at the optic chiasm. Identifying gene programs on how ipsi/contra RGC diversity arises reveals how decussating systems such as the binocular circuit are established. Such information is critical for driving stem cells into RGCs for replacement therapy and directing axon regeneration in injured and degenerating visual pathways.

项目总结 来自每只眼睛的视网膜神经节细胞(RGC)将它们的轴突本身和对侧延伸到大脑，以 建立双目视觉电路。研资局预测缺乏适当的侧面性正在削弱 遗传性疾病，如白化病，与视网膜色素上皮(RPE)色素减少有关 导致视神经错位，从而改变了立体视觉。这笔赠款的进展包括确定 IPSI和对应性RGC的规范和分化的转录调控因子.演示 腹侧睫状缘区(CMZ)作为IPSI RGC的来源；细胞周期调节因子Cyclin D2在 腹侧CMZ；以及IPSI RGC投射对Cyclin D2的依赖性。在白化视网膜中，这些过程 和RPE的细胞完整性被破坏，使白化病成为 建议进行的研究。 在这里，我们探索了CMZ在神经视网膜发育和RGC规范中的新角色，并试图 发现潜在的信号网络，以建立适当的RGC连接。我们建议将研究结合起来 用CMZ、RPE和神经视网膜的转录片段研究神经发生和命运图谱，以获得机制 洞察白化动物RPE中黑色素的丢失如何导致细胞命运从IPSI转变为相反 双目环路的建立。为此，我们将确立Cyclin D2在细胞中的作用 周期，细胞分裂平面，以及从CMZ到神经视网膜的迁移(目标1)；研究 RPE和CMZ，从Wnt途径开始，作为调节RGC命运的途径(目标2)；确定 调控RGC细胞命运的转录网络通过执行CMZ、神经视网膜、 和RPE(目标3)。从头到尾，我们都会比较白化视网膜和色素视网膜。我们假设世界上的事件 CMZ控制神经发生时间，它指定RGC投射命运和RPE中提供的因素 指示CMZ和神经视网膜获得IPSI/对应性RGC命运。 意义：探索早期神经视网膜发育和解析神经发生和细胞命运的调节因素 从CMZ和RPE发出的信息提供了一个新的机制切入点，以揭开长期存在的谜团 色素和RPE影响视交叉的细胞命运和RGC轴突分离。识别基因 有关ipsi/对数RGC多样性如何产生的程序揭示了交叉系统(如双目电路)如何 都是成立的。这些信息对于推动干细胞进入视网膜节细胞进行替代治疗和指导 损伤和退化的视觉通路中的轴突再生。