Molecular Basis of Outer Retina Development and Repair

外视网膜发育和修复的分子基础

• 批准号: 10269817
• 负责人: Melanie A Samuel
• 金额: $ 11.87万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-10-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10269817
• 关键词: 3-Dimensional; 5' -AMP-activated protein kinase; Adult; Axon; Blindness; Cells; Cone; Defect; Development; Disease; Ensure; Event; Genes; Genetic Transcription; Image; Imaging Techniques; Interneurons; Knockout Mice; Light; Mediating; Molecular; Neurons; Pathway interactions; Phosphotransferases; Photoreceptors; Process; Protein-Serine-Threonine Kinases; Retina; Retinal Defect; Rod; STK11 gene; Signal Pathway; Signal Transduction; Synapses; Testing; Time; Vision; Visual; genetic analysis; interest; novel; premature; prevent; reconstruction; repaired; retinal neuron; visual information

PROJECT SUMMARY Vision begins when light detecting photoreceptors in the outer retina sense and respond to visual input and relay this information to interneurons. Both the development of these connections and their long-term integrity must be precisely regulated in space and time to ensure visual information is correctly relayed. Understanding the mechanisms that coordinate these events is central to understanding the basis of many visual diseases. We have identified that the serine-threonine kinase LKB1 is differentially required for the emergence and long- term fidelity of the outer retina. Our preliminary evidence suggests that deletion of LKB1 during development inhibits outer retina synaptic formation, while deletion in adulthood causes premature synaptic decline. We are therefore extremely interested to understand how LKB1 orchestrates these distinct processes at both the cellular and molecular level. Our preliminary evidence suggests that the cells and signaling pathways that initiate and regulate these events are independent. Adults require LKB1 signaling in rods to ensure the organization of their synapses, while development of the outer retina requires LKB1 driven extension of cone axons. Moreover, LKB1 signals through district pathways to mediate these events: in adults LKB1 controls synaptic fidelity through the AMP activated kinase AMKP, while LKB1 functions independently of AMPK in development. Our first Aim will determine the precise cellular requirement for LKB1 in the outer retina during development and adulthood using cell-specific knockout mice, live imaging, and single cell reconstruction. We will also interrogate the impact of outer retina defects on the molecular organization the synapses that reside there using 3D nanoscopic imaging techniques. Our second aim will determine the mechanism by which LKB1 functions using genetic analyses of LKB1-kinase pathways and cell-specific transcriptional approaches. Finally, we will test whether manipulating AMPK or other targets can prevent or reverse visual decline. These studies will lead to the identification of novel molecular pathways for manipulating retina circuits that may ultimately be useful for repairing them.

项目摘要 当光检测视网膜外感光体并响应视觉输入和 将此信息传递给中间神经元。这些联系的发展及其长期完整性 必须在空间和时间上进行精确调节，以确保视觉信息正确中继。理解 协调这些事件的机制对于理解许多视觉疾病的基础至关重要。 我们已经确定，丝氨酸 - 硫代激酶LKB1的出现需要差异 外视网膜的术语保真度。我们的初步证据表明，在开发过程中删除LKB1 抑制视网膜外突触形成，而成年后的缺失会导致早产。我们是 因此，非常有兴趣了解LKB1如何在这两个方面策划这些不同的过程 细胞和分子水平。我们的初步证据表明，细胞和信号通路 启动和规范这些事件是独立的。成人需要杆中的LKB1信号，以确保 组织突触的组织，而视网膜外部的发展需要LKB1驱动的锥体扩展 轴突。此外，LKB1通过地区途径进行调解这些事件的信号：在成年人中LKB1对照 通过AMP激活的激酶AMKP的突触保真度，而LKB1独立于AMPK发挥作用 发展。我们的第一个目标将确定外部视网膜中LKB1的精确细胞需求 使用细胞特异性基因敲除小鼠，实时成像和单细胞重建的发育和成年。我们 还将询问外部视网膜缺陷对分子组织的影响 那里使用3D纳米镜成像技术。我们的第二个目标将确定LKB1的机制 使用LKB1-激酶途径和细胞特异性转录方法的遗传分析的功能。最后， 我们将测试操纵AMPK或其他目标是否可以预防或逆转视觉下降。这些研究 将导致鉴定新的分子途径来操纵视网膜电路，最终可能是 可用于修理它们。