SNAP25b介导的神经支配调控多疣壁虎断尾再生的机制研究

The successful regeneration of tissue or organ requires precise innervation. However, it is still incompletely clarified how the innervation is regulated and what the releasing crucial factors are. It is generally acceptable that synaptic vesicles are important organelles to carry out this function. Our previous findings have shown that the presynaptic protein SNAP25b was able to promote axonal regeneration by regulation of vesicle secretion after gecko tail amputation, suggesting that this protein is involved in the innervations of regenerating tissues or organs. In the present project, we are going to examine the relations between expression of SNAP25b and axonal regeneration in the injured spinal cord following gecko tail amputation; by interference of SNAP25b expression in vivo, we try to examine its effects on axonal regeneration, the formation of WE, as well as the proliferation and differentiation of blastema cells. We also attempt to investigate the number and migration of vesicles by interference of SNAP25b expression in SH-SY5Y cells. By using protein-mass-spectrometric technology, we are going to analyze the innervation-related growth factors in response to SNAP25b transfection, following vesicles collection from the primary cultured neurons of gecko. On the basis of the findings, we further validate these factors in injured spinal cord. Finally, we will unveil how SNAP25b is regulated during the spinal cord regeneration. The investigation of SNAP25b in the innervations of tail regeneration will provide new clues for the successful regeneration of tissues or organs in the mammal adults.

组织或器官再生需要神经支配才能完成。但如何调控神经支配以及神经支配相关的物质基础目前仍不十分清楚。突触囊泡是完成这一功能的重要载体。我们前期工作发现，突触前膜特异性蛋白SNAP25b调节囊泡分泌，参与壁虎断尾神经轴突再生，提示与组织器官再生的神经支配有关。本课题拟采用壁虎断尾模型，检测损伤脊髓中SNAP25b的表达变化与再生轴突之间的相关性；在体干预SNAP25b表达，检测SNAP25b对轴突再生、受伤表皮形成、芽基细胞增殖分化的影响；采用神经元细胞模型，干预SNAP25b表达，观察其囊泡数量和迁移的影响；蛋白质谱技术分析壁虎原代皮层神经元囊泡中的再生相关因子，揭示SNAP25b介导神经支配的物质基础，并进行在体验证；分析断尾脊髓再生过程中调控SNAP25b表达的关键因子。通过对SNAP25b介导的神经支配调控多疣壁虎断尾再生的机制研究，为高等动物的组织器官再生提供重要参考。

组织或器官再生需要神经支配才能完成。但如何调控神经支配以及神经支配相关的物质基础目前仍不十分清楚。突触囊泡是完成这一功能的重要载体。我们发现，突触前膜特异性蛋白SNAP25b调节囊泡分泌，参与壁虎断尾神经轴突再生，提示与组织器官再生的神经支配有关。本课题拟采用壁虎断尾模型，发现损伤脊髓中SNAP25b的表达变化与再生轴突之间的时空相关性以及再生芽基组织中的表达分布；采用未分化PC12细胞模型，过表达SNAP25b，发现其能促进细胞分化及突起生长；原代培养多疣壁虎皮层神经元，过表达SNAP25b，并收集其囊泡，发现其分泌数量增多；微流体培养大鼠DRG并进行轴突横断，收集上述囊泡处理神经元，发现囊泡促进轴突再生，揭示SNAP25b介导的囊泡分泌是神经再生的物质基础。通过对SNAP25b介导的神经支配调控多疣壁虎断尾再生的机制研究，为高等动物的组织器官再生提供重要参考。