【摘要】 目的探究突触黏附分子NECL4对Ca²⁺-CaMKII-CDC42信号通路和树突棘数量的影响。方法以Necl4全身性敲除小鼠为研究对象,对8周龄的Necl4野生（WT）及敲除（KO）小鼠脑组织进行高尔基染色,对大脑皮质第Ⅱ/Ⅲ层椎体神经元二级树突上树突棘密度进行统计。通过Western blot和RT-qPCR实验对Ca²⁺-CaMKII信号通路及其下游信号分子的蛋白和mRNA表达水平进行检测,包括CDC42、Rac1、RhoA、H-Ras、ERK等信号蛋白。结果高尔基染色结果显示,Necl4敲除小鼠大脑皮质第Ⅱ/Ⅲ层椎体神经元二级树突上树突棘数量与野生型小鼠相比减少（P<0.01）。Western blot实验结果显示CaMKIIα（P<0.05）及phospho-CaMKIIα（Thr286）（P<0.05）在Necl4敲除小鼠中表达降低,CaMKIIα的下游信号分子CDC42的mRNA水平（P<0.05）和蛋白水平（P<0.05）在Necl4敲除小鼠中均表达降低。结论 NECL4通过Ca²⁺-CaMKII-CDC42信号通路对小鼠大脑皮质树突棘数量进行调控。更多还原

【Abstract】 Objective To investigate the effect of synaptic adhesion molecule nectin-like molecule 4（NECL4） on Ca²⁺-CaMKII-CDC42 signal pathway and the number of dendritic spines. Methods Using Necl4 knockout mice as the research animal, the brain tissues of 8-week-old Necl4 wild type（WT） and knockout（KO） mice were stained with Golgi staining, then the density of dendritic spines on the secondary dendrites in layer Ⅱ/Ⅲ of cerebral cortex neuron was examined. Western blot and RT-qPCR were used to detect the protein and mRNA expression of Ca²⁺-CaMKII signal pathway and downstream signal molecules, including CDC42, Rac1, RhoA, H-Ras, ERK andother signal proteins.Results The Golgi staining showed that the number of dendritic spines on the secondary dendrites of the cerebral cortex Ⅱ/Ⅲ layer neuron in Necl4 knockout mice was less than that in wild type mice（P<0.01）. The results of Western blot showed that the expression of CaMKIIα（P<0.05） and phospho-CaMKIIα（Thr286）（P<0.05） decreased in Necl4 knockout mice. The mRNA（P<0.05） and protein（P<0.05） levels of CDC42 were decreased in Necl4 knockout mice. Conclusions NECL4 regulates the number of dendritic spines in mouse cerebral cortex through Ca²⁺-CaMKII-CDC42 signal pathway.更多还原