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: 10652910
• 负责人: Melanie A Samuel
• 金额: $ 7.22万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10652910
• 关键词: 3-Dimensional; 5' -AMP-activated protein kinase; AMP-activated protein kinase kinase; Adult; Affect; Animals; Antibodies; Axon; Biochemical Genetics; Blindness; Cells; Clinical; Cone; Data; Defect; Development; Disease; Ensure; Event; Genes; Genetic Transcription; Goals; Growth; Image; Imaging Techniques; Individual; Interneurons; Knockout Mice; Light; Maintenance; Maps; Mediating; Molecular; Molecular Target; Mutant Strains Mice; Neurons; Outcome; Pathology; Pathway interactions; Phosphotransferases; Photoreceptors; Population; Process; Protein-Serine-Threonine Kinases; Reagent; Regulation; Research; Retina; Retinal Defect; Rod; STK11 gene; Shapes; Signal Pathway; Signal Transduction; Structural defect; Synapses; Techniques; Testing; Therapeutic; Time; Variant; Viral; Vision; Visual; Work; experimental study; genetic analysis; genetic approach; imaging approach; interest; mutant; neuron development; novel; premature; prevent; reconstruction; repaired; retinal neuron; synaptogenesis; transcriptome sequencing; 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 当外部视网膜中的光检测光感受器感知并响应视觉输入时， 将这些信息传递给中间神经元。这些联系的发展及其长期的完整性 必须在空间和时间上进行精确调节，以确保视觉信息得到正确传递。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-苏氨酸激酶LKB 1是不同的需要为出现和长期- 外部视网膜的长期保真度。我们的初步证据表明，在发育过程中LKB 1的缺失 抑制外层视网膜突触的形成，而成年期的缺失会导致突触过早衰退。我们 因此，非常有兴趣了解LKB 1如何在两个不同的过程中编排这些不同的过程， 细胞和分子水平。我们的初步证据表明， 发起和管理这些事件是独立。成年人需要LKB 1信号在杆，以确保 外视网膜的发育需要LKB 1驱动的视锥细胞的延伸， 轴突此外，LKB1信号通过区域途径介导这些事件：在成人中，LKB1对照 突触保真度通过AMP激活的激酶AMKP，而LKB 1的功能独立于AMPK， 发展我们的第一个目标是确定在视网膜外膜中LKB 1的精确细胞需求。 使用细胞特异性敲除小鼠、活体成像和单细胞重建进行发育和成年。我们 还将询问外层视网膜缺陷对分子组织的影响， 使用3D纳米成像技术。我们的第二个目标是确定LKB 1 使用LKB1-激酶途径的遗传分析和细胞特异性转录方法，最后， 我们将测试是否操纵AMPK或其他目标可以防止或逆转视力下降。这些研究 将导致识别操纵视网膜电路的新分子途径， 用于修复它们。