PNN细胞外基质对再造神经组织中神经突触可塑性的影响及其分子机制研究

The latest research suggests that extracellular matrix (ECM) of perineuronal nets (PNN) play an very important role in regulating the formation, maturation, maintenance and function of synapses. Emerging data support the view that ECM aberrations are likely to contribute to imbalanced synaptic function in epilepsy, Alzheimer's disease and other neurodegenerative disorders, mental retardation and schizophrenia. The project aims to investigate the effect of different ECM on the synaptic plasticity through microprinting techique and screen the PNN　ECM gredients possessing the ability to promote the formation and maturation of synapse. At the same time, utilizing primary neural cells and neural stem cells as seeding cells and collagen type I gel combined with different ECM as a scaffolds, we will reconstruct three-dimensional neural tissue in vitro through tissue engineering technique, study the condition and regularity of primary neural cells and neural stem cells self-assembly into neural tissue and further development in three-dimensional culture condition. On the basis of previous results, we will study the regularity of the influence of PNN ECM on the structure and function of neurons such as ion channel and cytoskeleton protein in reconstructed neural tissue and further explore their molecular mechnisms through histology and morphology, molecular biology, electrophysioloy etc. The project is significant to bright light on the pathologic mechanism of nervous stystem disease such as epilepsy and neurodegenerative disorders. Moreover, the acquired achievements will provide important data for the investigation of neural tissue engineering and further enhance the quality of reconstructed three-dimensional neural tissue.

最新研究表明，PNN细胞外基质对神经突触形成、成熟与稳定发挥至关重要的作用，与癫痫、神经退行性疾病以及精神分裂症等疾病发生密切相关。本项目将摸索并建立微印刷制备体系和技术平台；以此为手段，研究不同PNN细胞外基质对神经突触可塑性的影响，筛选促进神经突触形成与成熟的细胞外基质成分；利用组织工程技术，以原代神经细胞及神经干细胞为种子细胞，以复合不同细胞外基质成分的I型胶原凝胶等为支架材料，进行神经组织体外三维再造研究，摸索并掌握神经细胞在三维条件下自组装形成神经组织及进一步发育的条件和规律；在此基础上，以再造神经组织为模型，研究PNN细胞外基质对再造神经组织中神经元离子通道与骨架蛋白等结构与功能的影响，并深入探索其影响神经突触可塑性的分子机制。本项目研究对揭示癫痫、神经退行性疾病等神经疾病的发病机理具有重要意义，且所获成果将为神经组织工程研究提供重要实验数据，进而提高体外再造三维神经组织质量。

本项目摸索并建立微印刷制备体系和技术平台，并以此为手段，研究了PNN细胞外基质对突触可塑性的影响；并利用组织工程技术，以原代神经细胞以及神经干细胞为种子细胞，以复合促进神经突触形成与成熟的PNN细胞外基质成分的Ⅰ型胶原凝胶等为支架材料，进行神经组织体外三维再造研究，优化并建立神经组织三维再造体系；在此基础上，研究电磁刺激下的小脑切片兴奋性与功能连接性的频率依赖性调制，深入解析了刺激后的功能连接性；研究了植入式电极表面改性，具有良好的应用潜能。