荧光探针逆行标记投射至小脑的前庭内侧核神经元及其活性评价  

Identification method and electrophysiological evaluation of cerebellum-projecting anteromedial nucleus neurons by retrograded fluorescence probe

作　　者：周荣超[1] 胡娟娟[1] 张秀娟[1] 朱云[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院耳鼻咽喉头颈外科,武汉430022

出　　处：《中华实验外科杂志》2017年第8期1333-1335,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81600805）

摘　　要：目的 探索甄别投射至小脑的前庭内侧核（MVN）神经元的方法,并对其活性进行评价.方法 应用在体荧光探针逆行标记投射至小脑的MVN神经元,应用中性红染色鉴定细胞存活时间,应用全细胞膜片钳技术记录其基础电生理特性,并与未经探针标记的MVN神经元进行比较.结果 投射至小脑的MVN神经元（CPMVNs）与未经探针标记的SD大鼠MVNs在离体1、2、3、4、5h的条件下存活时间比较差异无统计学意义;自发性放电频率及膜电位的电生理比较差异无统计学意义;两组神经元单突触激发的兴奋性突触后电流（eEPSCs）幅值分别为（267.100±7.563）pA（n=18）和（266.900±6.730）pA（n=25）,组间比较差异无统计学意义（P=0.962）;两组神经元自发性抑制性突触后电流（sIPSCs）幅值及频率分别为（19.320±1.768）pA及（2.936±0.512）Hz（n=21）,（18.360±1.700）pA及（2.871±0.454）Hz（n=27）,两组sIPSCs在电流幅值（P=0.207）及频率（P=0.755）比较差异均无统计学意义.结论 应用在体荧光探针逆行标记技术甄别活性良好的CPMVNs,可为进一步研究前庭小脑通路上突触信号传导机制提供特异性实验对象.Objective To explore sufficient ways to identify the anteromedial nucleus （MVNs） neurons that project to the cerebellum and evaluate their activity.Methods Retrograde fluorescence probe injection techniques were in vivo applicated to MVNs neurons that project to the cerebellum,neutral red staining was used to determine cell survival time,electrophysiological characteristics by whole cell patch clamp technique were recorded between retrograde labeled anteromedial nucleus neurons projecting to the cerebellum （CPMVNs） and unlabeled MVNs.Results Typical electrophysiological characteristics of retrograde labeled CPMVNs,including spontaneous firing frequency,membrane potential,excitatory presynaptic glutamate afferent and inhibitory postsynaptic gamma aminobutyric acid （GABA） ergic afferent showed no statistical significance,compared with unlabeled MVNs.The amplitudes of monosynaptically evoked excitatory postsynaptic currents （eEPSCs） were respectively （267.100±7.563） pA （n=18）,（266.900±6.730） pA （n=25）,P=0.962.The current amplitude and frequency of spontaneous inhibitory postsynaptic currents （sIPSCs） were respectively（19.320±1.768）pA and （2.936±0.512） Hz （n=21）,（18.360±1.700） pA and （2.871±0.454） Hz （n=27）,with P values were 0.207,0.755 separately,which showed no statistical significance.Conclusion Vivo application of retrograde fluorescence probe injection techniques helps to identify good CPMVNs,which are specific subjectsfor further study on synaptic transmission mechanism of vestibular-cerebellum pathway.

关 键 词：前庭内侧核 电生理 逆行标记 脑片 突触传递 

分 类 号：R338[医药卫生—人体生理学]