摘要
目的:通过激活未定带(ZI)γ-氨基丁酸(GABA)能神经元,观察脂多糖(LPS)处理后小鼠运动能力损伤的缓解情况,探讨GABA能神经元在中枢神经系统感染模型小鼠运动功能损伤过程中的作用.方法:利用腹膜腔注射LPS方法制备C57BU6小鼠中枢神经系统感染模型,利用旷场实验与转轮实验检测小鼠运动功能的变化,免疫荧光染色观察小鼠ZI神经元c-Fos和离子钙结合适配器分子1(Iba-1)的表达.利用立体定位技术将rAAV-DIO-hM3Dq-mCherry注射至成年雄性VGAT-Cre转基因小鼠双侧ZI区,等待3周病毒表达充分后,进行腹腔注射LPS制备神经炎症感染模型,行为检测30 min前腹腔注射N-氧化氯氮平(CNO)激活GABA能神经元,利用旷场实验与转轮实验检测小鼠运动功能的变化.结果:旷场实验与转轮实验结果显示,与对照组相比,LPS组小鼠活动度显著降低(P＜0.05),小鼠ZI内c-Fos阳性蛋白表达显著下调,同时Iba-1阳性蛋白表达显著增加;并且化学遗传结果显示,与LPS组相比,激活小鼠ZI部位GABA能神经元后LPS小鼠活动能力显著增强(P＜0.05).结论:神经炎症状态小鼠ZI部位GABA能神经元活性显著降低,运动能力受损,化学遗传激活ZI部位GABA能神经元显著改善了炎症状态小鼠的运动能力.
Abstract
Objective:By activating gamma-aminobutyric acid(GABA)neurons in zona incerta(ZI),we observed the alleviation of motor ability impairment in mice treated with lipopolysaccharide(LPS),and investigate the function of GABA neurons in the process of motor function impairment in mice with neuroinflammation.Methods:The central nervous system infection model of C57BU6 mice was eatablished by intraperitoneal injection of LPS.The motor function were detected by open field test and running wheel test in mice.The expression of c-Fos in ZI neurons and ionized calci-um binding adaptor molecule 1(Iba-1)in glial cells were detected by immunofluorescence staining.rAAV-DIO-hM3Dq-mCherry was bilaterally injected into ZI of adult male VGAT-Cre mice through the stereotaxic technique.After three weeks with the virus sufficiently expressing in the neurons,the neuroinflammation model was made by intrabitoneal injection of LPS,and GABAergic neurons were activated by intrabitoneal injection of N-oxyclozapine(CNO)30 min before behavioral tests.The motor ability in mice were detected by open field test and running wheel test.Results:The results of open field experiment and wheel experiment showed that the activity of mice in LPS group was significantly decreased(P＜0.05)compared with the control group.The expression of ZI c-Fos positive protein in the brain of mice was significantly down-regulated with Iba-1 positive protein up-regulating in the ZI.The chemogenetic results showed that the activity of LPS mice was relieved with ZI GABAergic neurons being activated(P＜0.05).Conclusion:The activity of ZI GABAergic neurons was significantly decreased and the motor ability was impaired in mice with neuroin-flammation in the brain.Chemogenetic activation of ZI GABAergic neurons significantly improved the motor ability of inflammatory mice.
基金项目
国家自然科学基金(81870852)
国家自然科学基金(82171199)
江苏省大学生创新训练课题(202210313037Z)
NETL
NSTL
万方数据