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目的:建立光遗传-在体多通道光电极记录系统,用其对直接通路中型多棘神经元(D1-MSN)的活动模式进行在体记录。方法:首先向D1-Cre转基因小鼠纹状体背外侧注射携带光敏阳离子通道channelrhodopsin-2(ChR2)基因的腺相关病毒,使ChR2在D1-MSN上通过Cre重组酶的同源重组而特异性表达。之后通过光刺激和电生理记录相结合的方式在体记录D1-MSN的活动模式。结果:D1-Cre小鼠纹状体在注射病毒后通过荧光显微镜观察,可发现明显的荧光信号,证明病毒正常表达。通过光遗传-在体多通道光电极记录系统,本研究成功地在纹状体内用光刺激诱发了MSN的电活动。通过对记录的电信号进行数据分析,证明了光刺激诱发的电信号确实来自于D1-MSN,成功地对D1-MSN活动模式进行了在体记录。结论:结果提示光遗传-在体多通道光电极记录系统是一种对纹状体D1-MSN电活动模式记录的可选新方法。
OBJECTIVE: To establish a phototransmission-in-vivo multi-channel optoelectrode recording system for on-body recording of the activity pattern of direct pathway mesial spine neurons (D1-MSN). METHODS: The adeno-associated virus carrying the channelrhodopsin-2 (ChR2) -positive cation channel of D1-Cre transgenic mice was dorsolateral injected into the striatum of D1-Cre transgenic mice to make ChR2 specific by homologous recombination of Cre recombinase on D1-MSN expression. The patterns of D1-MSN activity were then recorded in vivo by a combination of photo-stimulation and electrophysiological recording. Results: The striatum of D1-Cre mice was observed by fluorescence microscopy after virus injection. Fluorescent signal was observed and the virus was normally expressed. Through phototransmission-in-vivo multi-channel photoelectrode recording system, the present study successfully stimulated the electrical activity of MSN with light stimulation in the striatum. Through the data analysis of the recorded electrical signals, it was proved that the electrical signals induced by photo-stimulation did come from D1-MSN and the on-body recording of D1-MSN activity patterns was successfully performed. CONCLUSIONS: The results suggest that the mesogen-in-vivo multi-channel optoelectrode recording system is an alternative new method for recording electrical activity patterns of the D1-MSN striatum.