胚胎发育期五羟色胺转运体基因沉默对皮层锥体神经元发育的影响

Serotonin transporter (Sert) is the key modulator for the homeostasis of 5-HT and the development of serotoninergic neurons. It has been proved that the imbalance of the homeostasis of 5-HT neurons is one of the main pathological mechanisms for psychiatric disorders, such as, autism and schizophrenia. We hypothesize that the increase of 5-HT level caused by Sert gene regulation would affect the cortical neuron development. The new developed in-utero electroporation (IUE) technique provides an easy-to-use, safe, high-efficiency in-vivo gene-transfer way to study the gene function in central nerve system. Thus, this project aims to construct the first combined IUE and RNA interference (RNAi) platform, through which to transfer the Sert shRNA - GFP plasmid to the cerebral cortex of E14.5 mouse embryo. This project will observe the gene silence effects of Sert on the migration of pyramid neuron in cerebral cortex, as well as the appearance of the pyramid neuron in different develop stage. These observations will help to explain the developmental dysregulation of serotonin homeostasis contributes to increased vulnerability to psychiatric disorders, such as, autism. This project will provide a new method to generate the autism animal model, while the IUE-RNAi platform will be useful for novel drug development as well as the clinical treatment.

5-羟色胺转运体（Sert）是调节5-羟色胺（5-HT）内稳态及其神经元发育的关键基因。研究证明5-羟色胺能神经元发育稳态失衡是导致自闭症及精神分裂症等精神类疾病的重要病理机制之一。我们拟通过调节Sert表达改变皮层5-HT水平，进而影响皮层神经元的发育。新兴的子宫内电穿孔技术IUE提供了方便、安全、高效的活体基因转染方法，研究基因在中枢神经系统发育中的功能。本课题首次建立IUE结合RNA干扰（RNAi）这一先进技术平台，将构建的Sert shRNA-GFP质粒通过IUE导入E14.5鼠胚大脑皮层，观察鼠胚皮层Sert基因沉默对其锥体神经元迁移的影响，同时观察其对不同发育期鼠胚锥体神经元形态的影响，从而验证5-HT能神经系统发育障碍在自闭症中的病理机制。通过本课题研究将为建立神经发育障碍所致自闭动物模型提供新的思路，建立的IUE-RNAi技术平台也将为新药研发及临床治疗开辟新的途径。