本课题希望精确、定量地揭示AMPA受体剪接亚型（flip/flop）对突触传递功能和可塑性的贡献。AMPA受体介导了脑内最主要的快速兴奋性神经传递。在以前的工作（Neuron，2009）中，申请者和同事发展了单细胞基因敲除法，阐明了脑内椎体神经元突触膜上的AMPA受体主要由GluA1/A2异源四聚体组成。然而，每种AMPA受体亚基由于mRNA剪接造成至少flip/flop两种亚型，使突触AMPA受体更加多样化。最近的报道表明受体编码mRNA的剪接受到神经活动的调节。那么如何从功能上区分突触上flip和flop类型的AMPA受体呢？本课题将借助新近发展的CRISPR-Cas9基因工程技术，特异性地剔除AMPA受体flip和flop的编码外显子，用遗传学方法分离出纯净的AMPA受体剪接亚型，并结合双电极电生理技术精确研究它们的突触功能，以深入理解突触受体可塑性变化受神经活动调节的机理。

AMPA-type glutamate receptors (AMPARs) mediate the most fast excitatory neurotransmission in brain. The AMPARs on synaptic membrane play critical roles in regulating the transmission efficiency and synaptic plasticity. In order to pinpoint the molecular composition of synaptic AMPARs, we have previously developed single-cell genetics and shown that the synaptic AMPARs in pyramidal neurons are made of GluA1/A2 heteromers (Neuron, 2009). However, each AMPAR subunits could form flip/flop isoforms resulting from alternative mRNA splicing. Recent reports demonstrate that neural activity shaped AMPAR mRNA splicing, indicating change of synaptic AMPAR components and functions. To explore the precise roles that flip/flop type receptors play in synapse, we propose to isolate the AMPAR isoforms and study their synaptic function by targeting the specific DNA segments (exons) encoding the flop and flip isoforms using CRISPR-Cas9 techniques. This study will hopefully reveal the underlying mechanisms for synaptic plasticity induced by neural activity.