中枢神经再生受多条信号通路调控，抑制Rho/ROCK信号通路能促进损伤中枢神经的轴突再生和功能恢复。14-3-3是调控信号通路的整合子，能与RhoA-GEF相互作用。 然而，14-3-3能否通过调控RhoA/ROCK信号通路影响中枢神经再生及其调节机制的问题，目前尚不清楚。我们前期研究发现14-3-3ε、RhoA和ROCK主要表达在涡虫中枢神经，并参与中枢神经再生。本课题拟从克隆在进化上占据重要地位的涡虫14-3-3基因入手，利用原位杂交、免疫组化和RNAi技术，研究14-3-3在涡虫中枢神经再生中的分布、表达及功能；通过免疫共沉淀筛选出14-3-3相互作用蛋白并分析其与涡虫中枢神经再生的关系；体内干预14-3-3及其相互作用蛋白表达，检测其对涡虫再生神经和RhoA/ROCK信号通路的影响，明确其功能和调控机制。该研究将为动物中枢神经再生提供理论依据，进而为脊椎动物再生机制研究提供新思路。

The regeneration of central nervous system (CNS) is regulated by the multiple signaling pathways. The inhibition of Rho/ROCK pathway can promote axon regeneration and functional recovery in the injured CNS. 14-3-3 proteins can interact with RhoA-GEF as a integrator in the signaling pathway. However, whether 14-3-3 affects the regeneration of CNS through regulating RhoA/ROCK signaling pathway and its regulating mechanism is unclear. Our previous studies revealed that 14-3-3 ε, RhoA and ROCK expressed in CNS, and involved in the process of the regeneration of planarian CNS. To address the expression pattern and function of 14-3-3 in the CNS regeneration, Firstly，we will clone the sequence of 14-3-3 genes in the planarian which occupies an important position in the evolution. Next, the location, expression patterns and function of 14-3-3 during planarian regeneration will be confirmed by immunofluorescence, in situ hybridization and RNAi. Thirdly, the interaction proteins with 14-3-3 are screened using co-immunoprecipitation, and analyze its relationship with the regeneration of planarian CNS. Finally, in order to confirm the function and mechanism of 14-3-3 regulated RhoA/ROCK pathway during the process of regeneration, we will further examine the effects on the planarian CNS regeneration and RhoA/ROCK pathway by interference of 14-3-3 and its interaction proteins. In conclusion, the function and molecular mechanisms of 14-3-3 regulated RhoA/ROCK pathway on planarian CNS regeneration will be illuminated by the aid of the knowledge of neurosciences and developmental biology. The project will provide the evidence of further researching the CNS regeneration of animal, and even provide new clue for the research on the mechanism of regeneration in vertebrates.