单细胞水平干预微小RNA调控斑马鱼毛特纳运动神经元轴突再生

脊髓损伤是一种中枢神经系统损伤疾病，会致损伤发生的脊髓节段及以下自主神经功能障碍。以往脊髓损伤的研究建立了多种轴突损伤模型，发现了一些轴突损伤再生的机制和调控手段，阐述了神经再生的内源性因素和外在影响因素。但是神经束中少量神经元轴突可以越过胶质疤痕再生说明其内源性因素相当重要，深入阐明这一机制需要建立中枢神经单细胞轴突损伤和再生模型。据此我们结合斑马鱼在位活体成像的优势及前期已成功建立的单细胞准确高效的干预手段，建立单个毛特纳轴突毁损再生模型，并对调控轴突再生的若干内源性因素进行深入探究。同时，微小RNA作为重要的非编码序列，在神经元分化，尤其是轴突发生延展过程中起到了重要的调控作用。考虑轴突再生过程与发生过程的相似性，本项目拟通过毛特纳单细胞水平干预基因表达调控、活体在位成像等，深入探索微小RNA对于轴突再生的调控作用及其机制，并论证部分微小RNA用于轴突损伤后再生的潜在有效性。

Spinal cord injury is a common central nervous system injury disease, which will lead to the dysfunction of autonomic nerve in certain spinal cord segment and downstream segment. With the research of spinal cord injury, a variety of neural axon injury models have been established, from which some mechanisms and regulatory means of axon regeneration have been found, including Waller's degeneration, inflammatory reaction, glial scar aggregation and some other endogenous factors. However, axons of some neurons in the nerve bundle can cross the glial scar to show the important role of endogenous factors. Based on the advantages of zebrafish in vivo imaging and the accuracy and efficiency of single cell intervention, we established the Mauthner axon degeneration and regeneration model, and carried out in-depth research on several endogenous factors regulating axon regeneration..At the same time, microRNA, as an important gene family and noncoding sequence, plays an important role in the process of neuron differentiation, especially axon extension. Considering the similarity between axon regeneration process and extension process, this project intends to deeply understand the regulatory role of microRNA in axon regeneration, explore the mechanism of regulation, and demonstrate the effectiveness and possibility of some microRNAs used in the treatment of axon injury through single cell level intervention in gene expression regulation, in-situ imaging and other technologies.