嗅觉诱导的胡须摆动联合式学习记忆维持和消退的细胞机制研究
发布时间：2018-08-16 13:06
【摘要】：目的:研究嗅觉诱导的胡须摆动联合式学习记忆维持和消退与桶状皮层和运动皮层神经元功能变化的关系。方法:1.造模:选择出生20天体重大小一致的C57 Thy1-YFP小鼠,利用嗅觉诱导的胡须摆动装置,对小鼠进行乙酸丁酯嗅觉刺激和5Hz机械摆动胡须刺激的联合式训练。每天训练5次,每次训练20s。连续训练10天后暂停训练一周,一周后再次联合式训练一天,整个过程动态监测小鼠胡须摆动轨迹的变化。对照组不作联合式训练。2.行为监测:在每天联合式训练结束后,通过只给予乙酸丁酯嗅觉刺激诱导胡须摆动,利用高清数字摄像机拍摄小鼠胡须摆动视频,用图像分析软件分析胡须摆动频率、角度、潜伏期的变化。3.膜片钳实验:根据实验目的的不同,选择不同训练天数的小鼠作为电生理实验对象。自第一天训练开始分别选择训练第10天、第17天(消退训练第7天)、第18天(消退后再次联合式训练1天)以及对照组小鼠作为实验对象。利用膜片钳技术检测桶状皮层和运动皮层谷氨酸能神经元功能的变化。结果:(1)嗅觉和胡须刺激联合式训练10天后,嗅觉诱导的胡须摆动联合式学习记忆模型建立;模型建立后给予不配对刺激消退训练7天后,联合式学习记忆消退;消退后再次配对训练1天,联合式学习记忆重建:与对照组相比,联合式训练10天后实验组小鼠胡须摆动频率、角度、回缩时间明显增加(p0.01);与联合式训练第10天相比,消退训练7天后,小鼠胡须摆动频率、角度、回缩时间明显减小(p0.01);与消退训练7天后相比,再次给予联合式训练1天后,小鼠胡须摆动频率、角度、回缩时间明显增加(p0.01)。(2)在联合式记忆建立、消退、重建过程中,桶状皮层神经元功能持续上调:与对照组相比,实验组小鼠联合式训练10天后桶状皮层神经元编码动作电位数量增多(p0.01)、自发性兴奋性突触后电流(spontaneous excitatory postsynaptic current,s EPSC)发放频率和幅度增大(p0.01)、自发性抑制性突触后电流(spontaneous inhibitory postsynaptic current,s IPSC)发放频率和幅度减小(p0.01)。与联合式训练第10天相比,消退训练7天后桶状皮层在相同的刺激强度下产生的动作电位数量、s EPSC发放频率和幅度、s IPSC发放频率和幅度都没有统计学差异;与消退训练7天相比,再次联合式训练1天后桶状皮层神经元编码动作电位数量、s EPSC发放频率和幅度、s IPSC发放频率和幅度都没有统计学差异。(3)运动皮层神经元在联合式记忆建立时功能上调,在联合式学习记忆消退后神经元功能下调,在联合式学习记忆重建后神经元功能再次上调:与对照组相比,实验组小鼠联合式训练10天后运动皮层神经元编码动作电位数量增多(p0.01),s EPSC发放频率和幅度增大(p0.01),自发性抑制性突触后电流s IPSC发放频率和幅度减小(p0.01);与联合式训练第10天相比,消退训练7天后运动皮层在相同的刺激强度下产生的动作电位数量减少(p0.01)、s EPSC发放频率和幅度减小(p0.01)、s IPSC发放频率和幅度增加(p0.01);与消退训练7天相比,再次联合式训练1天后运动皮层神经元编码动作电位数量增加(p0.01)、s EPSC发放频率和幅度增大(p0.01)、s IPSC发放频率和幅度减小(p0.01)。结论:1.嗅觉诱导的胡须摆动联合式学习记忆维持与桶状皮层谷氨酸神经元功能上调有关。2.嗅觉诱导的胡须摆动联合式学习记忆消退与运动皮层谷氨酸能神经元功能下调有关。
[Abstract]:Objective: To study the relationship between olfactory-induced whisker oscillation and functional changes of barrel-like cortex and motor cortex neurons. Five times a day, 20 seconds each time. After 10 days of continuous training, the training was suspended for one week. After one week, the whole process of combined training was dynamically monitored. The control group did not do combined training. 2. Behavior monitoring: After the end of the daily combined training, only to give. Mouse's whisker oscillation was induced by butyl acetate olfactory stimulation. Mouse's whisker oscillation was videotaped by high-definition digital video camera. Mouse's whisker oscillation frequency, angle and latency were analyzed by image analysis software. 3. Patch clamp test: Mouse with different training days were selected as electrophysiological subjects according to different experimental purposes. After 10 days of training, 17 days (7 days of regressive training), 18 days (1 day of combined training after regressive training) and control group mice were selected as subjects. The changes of glutamatergic neurons in barrel cortex and motor cortex were detected by patch clamp technique. The combined learning and memory model of sensory-induced whisker swing was established; 7 days after the model was established, the combined learning and memory subsided after the unpaired stimulus subsidence training; 1 day after the subsidence, the combined learning and memory reconstruction was performed after the matching training: compared with the control group, the frequency, angle and retraction time of the whisker swing in the experimental group were 10 days after the combined training. Compared with the 10th day of combined training, the frequency, angle and retraction time of the whiskers of the mice significantly decreased after 7 days of regression training (p0.01); compared with the 7th day of regression training, the frequency, angle and retraction time of the whiskers of the mice significantly increased after 1 day of combined training (p0.01). In the process of reconstruction, the function of barrel cortical neurons was continuously up-regulated: compared with the control group, the number of barrel cortical neurons coded action potential increased (p0.01), spontaneous excitatory postsynaptic current (s EPSC) frequency and amplitude increased (p0.01) and spontaneous excitatory postsynaptic current (s EPSC) after 10 days of combined training in the experimental group. The frequency and amplitude of spontaneous inhibitory postsynaptic current (s IPSC) were decreased (p0.01). Compared with the 10th day of combined training, the number of action potentials, the frequency and amplitude of s EPSC, the frequency and amplitude of s IPSC, and the amplitude of action potentials produced by barrel cortex at the same stimulus intensity after 7 days of regression training were not found. There was no significant difference in the number of coding action potentials, frequency and amplitude of s EPSC firing, and frequency and amplitude of s IPSC firing in the barrel cortex neurons after 1 day of combined training compared with 7 days of regressive training. (3) Motor cortex neurons were up-regulated in the establishment of combined memory, and the neurons were up-regulated after combined learning and memory regression. Metafunction was down-regulated and neuronal function was up-regulated again after combined learning and memory reconstruction: Compared with the control group, the number of coding action potentials (p0.01), the frequency and amplitude of s EPSC firing increased (p0.01) and the frequency and amplitude of s IPSC firing decreased in the experimental group after 10 days of combined training. Compared with the 10 th day of combined training, the number of action potentials produced by motor cortex under the same stimulation intensity decreased (p0.01), the frequency and amplitude of s EPSC decreased (p0.01), and the frequency and amplitude of s IPSC increased (p0.01) after 7 days of combined training. Compared with the 7 th day of combined training, the motor cortex nerve after 1 day of combined training increased. The number of meta-coding action potential increased (p0.01), the frequency and amplitude of s EPSC increased (p0.01), and the frequency and amplitude of s IPSC decreased (p0.01). Conclusion: 1. Olfactory-induced whisker oscillation associated with the maintenance of learning and memory is related to the up-regulation of barrel cortex glutamate neurons. 2. Down regulation of glutamate neurons in motor cortex.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R338
本文编号：2186070
[Abstract]:Objective: To study the relationship between olfactory-induced whisker oscillation and functional changes of barrel-like cortex and motor cortex neurons. Five times a day, 20 seconds each time. After 10 days of continuous training, the training was suspended for one week. After one week, the whole process of combined training was dynamically monitored. The control group did not do combined training. 2. Behavior monitoring: After the end of the daily combined training, only to give. Mouse's whisker oscillation was induced by butyl acetate olfactory stimulation. Mouse's whisker oscillation was videotaped by high-definition digital video camera. Mouse's whisker oscillation frequency, angle and latency were analyzed by image analysis software. 3. Patch clamp test: Mouse with different training days were selected as electrophysiological subjects according to different experimental purposes. After 10 days of training, 17 days (7 days of regressive training), 18 days (1 day of combined training after regressive training) and control group mice were selected as subjects. The changes of glutamatergic neurons in barrel cortex and motor cortex were detected by patch clamp technique. The combined learning and memory model of sensory-induced whisker swing was established; 7 days after the model was established, the combined learning and memory subsided after the unpaired stimulus subsidence training; 1 day after the subsidence, the combined learning and memory reconstruction was performed after the matching training: compared with the control group, the frequency, angle and retraction time of the whisker swing in the experimental group were 10 days after the combined training. Compared with the 10th day of combined training, the frequency, angle and retraction time of the whiskers of the mice significantly decreased after 7 days of regression training (p0.01); compared with the 7th day of regression training, the frequency, angle and retraction time of the whiskers of the mice significantly increased after 1 day of combined training (p0.01). In the process of reconstruction, the function of barrel cortical neurons was continuously up-regulated: compared with the control group, the number of barrel cortical neurons coded action potential increased (p0.01), spontaneous excitatory postsynaptic current (s EPSC) frequency and amplitude increased (p0.01) and spontaneous excitatory postsynaptic current (s EPSC) after 10 days of combined training in the experimental group. The frequency and amplitude of spontaneous inhibitory postsynaptic current (s IPSC) were decreased (p0.01). Compared with the 10th day of combined training, the number of action potentials, the frequency and amplitude of s EPSC, the frequency and amplitude of s IPSC, and the amplitude of action potentials produced by barrel cortex at the same stimulus intensity after 7 days of regression training were not found. There was no significant difference in the number of coding action potentials, frequency and amplitude of s EPSC firing, and frequency and amplitude of s IPSC firing in the barrel cortex neurons after 1 day of combined training compared with 7 days of regressive training. (3) Motor cortex neurons were up-regulated in the establishment of combined memory, and the neurons were up-regulated after combined learning and memory regression. Metafunction was down-regulated and neuronal function was up-regulated again after combined learning and memory reconstruction: Compared with the control group, the number of coding action potentials (p0.01), the frequency and amplitude of s EPSC firing increased (p0.01) and the frequency and amplitude of s IPSC firing decreased in the experimental group after 10 days of combined training. Compared with the 10 th day of combined training, the number of action potentials produced by motor cortex under the same stimulation intensity decreased (p0.01), the frequency and amplitude of s EPSC decreased (p0.01), and the frequency and amplitude of s IPSC increased (p0.01) after 7 days of combined training. Compared with the 7 th day of combined training, the motor cortex nerve after 1 day of combined training increased. The number of meta-coding action potential increased (p0.01), the frequency and amplitude of s EPSC increased (p0.01), and the frequency and amplitude of s IPSC decreased (p0.01). Conclusion: 1. Olfactory-induced whisker oscillation associated with the maintenance of learning and memory is related to the up-regulation of barrel cortex glutamate neurons. 2. Down regulation of glutamate neurons in motor cortex.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R338
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