Diversity Supplement (Pilar Andrade) for Molecular Basis of Outer Retina Development and Repair

项目总结 当外部视网膜中的光感受器感觉到视觉输入并对其做出反应时，视觉开始 将这一信息传递给中间神经元。无论是这些联系的发展还是它们的长期完整性 必须在空间和时间上进行精确调控，以确保正确传递视觉信息。理解 协调这些事件的机制对于理解许多视觉疾病的基础是至关重要的。 我们已经证实，丝氨酸-苏氨酸激酶LKB1在细胞的出现和长期存活过程中起着不同的作用。 外部视网膜的术语保真度。我们的初步证据表明，在发育过程中LKB1的缺失 抑制视网膜外部突触的形成，而成年期的缺失会导致突触过早下降。我们是 因此，非常有兴趣了解LKB1如何在 细胞和分子水平。我们的初步证据表明， 这些事件的发起和调控是独立的。成年人需要杆状体内的LKB1信号来确保 它们的突触的组织，而外视网膜的发展需要LKB1驱动的锥体延伸 轴突。此外，LKB1信号通过区域通路来调节这些事件：在成人中，LKB1控制 突触的保真度是通过AMP激活的AMKP，而LKB1的功能不依赖AMPK。 发展。我们的第一个目标是确定外视网膜对LKB1的精确细胞需求 利用细胞特异性基因敲除小鼠、活体成像和单细胞重建来发育和成年期。我们 还将询问视网膜外部缺陷对驻留的突触的分子组织的影响 在那里使用了3D纳米成像技术。我们的第二个目标将决定LKB1 使用LKB1-激酶通路的遗传分析和细胞特异性转录方法的功能。最后， 我们将测试操纵AMPK或其他靶点是否可以防止或扭转视觉衰退。这些研究 将导致识别新的分子通路，以操纵视网膜电路，最终可能是 对修理它们很有用。

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.