Drosophila Laser Axotomy Injury Model to Investigate RNA Repair and Splicing in Axon Regeneration.
Drosophila Laser Axotomy Injury Model to Investigate RNA Repair and Splicing in Axon Regeneration.
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DOI：
10.1007/978-1-0716-3012-9_22
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发表时间：
2023
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成熟神经元的轴突再生能力有限，常常导致中枢神经系统（CNS）损伤后功能恢复不足。为了促进中枢神经系统神经修复，迫切需要了解再生机制，以开发有效的临床治疗方法。为此，我们开发了果蝇感觉神经元损伤模型和伴随的行为测定，以检查外周和中枢神经系统损伤后轴突再生能力和功能恢复。具体而言，我们使用双光子激光诱导轴突切断术，并进行实时成像以评估轴突再生，结合对热感受行为的分析作为功能恢复的读数。利用该模型，我们发现RNA 3′-末端磷酸环化酶（Rtca）作为RNA修复和剪接的调节剂，对损伤诱导的细胞应激作出反应，并在轴突断裂后阻碍轴突再生。在这里，我们描述了我们如何利用我们的果蝇模型来评估Rtca在神经再生过程中的作用。
The limited axon regeneration capacity of mature neurons often leads to insufficient functional recovery after damage to the central nervous system (CNS). To promote CNS nerve repair, there is an urgent need to understand the regeneration machinery in order to develop effective clinical therapies. To this aim, we developed a Drosophila sensory neuron injury model and the accompanying behavioral assay to examine axon regeneration competence and functional recovery after injury in the peripheral and central nervous systems. Specifically, we used a two-photon laser to induce axotomy and performed live imaging to assess axon regeneration, combined with the analysis of the thermonociceptive behavior as a readout of functional recovery. Using this model, we found that the RNA 3′-terminal phosphate cyclase (Rtca), which acts as a regulator for RNA repair and splicing, responds to injury-induced cellular stress and impedes axon regeneration after axon breakage. Here we describe how we utilize our Drosophila model to assess the role of Rtca during neuroregeneration.