發(fā)布時(shí)間：2018-08-16 13:06

【摘要】：目的:研究嗅覺(jué)誘導(dǎo)的胡須擺動(dòng)聯(lián)合式學(xué)習(xí)記憶維持和消退與桶狀皮層和運(yùn)動(dòng)皮層神經(jīng)元功能變化的關(guān)系。方法:1.造模:選擇出生20天體重大小一致的C57 Thy1-YFP小鼠,利用嗅覺(jué)誘導(dǎo)的胡須擺動(dòng)裝置,對(duì)小鼠進(jìn)行乙酸丁酯嗅覺(jué)刺激和5Hz機(jī)械擺動(dòng)胡須刺激的聯(lián)合式訓(xùn)練。每天訓(xùn)練5次,每次訓(xùn)練20s。連續(xù)訓(xùn)練10天后暫停訓(xùn)練一周,一周后再次聯(lián)合式訓(xùn)練一天,整個(gè)過(guò)程動(dòng)態(tài)監(jiān)測(cè)小鼠胡須擺動(dòng)軌跡的變化。對(duì)照組不作聯(lián)合式訓(xùn)練。2.行為監(jiān)測(cè):在每天聯(lián)合式訓(xùn)練結(jié)束后,通過(guò)只給予乙酸丁酯嗅覺(jué)刺激誘導(dǎo)胡須擺動(dòng),利用高清數(shù)字?jǐn)z像機(jī)拍攝小鼠胡須擺動(dòng)視頻,用圖像分析軟件分析胡須擺動(dòng)頻率、角度、潛伏期的變化。3.膜片鉗實(shí)驗(yàn):根據(jù)實(shí)驗(yàn)?zāi)康牡牟煌?選擇不同訓(xùn)練天數(shù)的小鼠作為電生理實(shí)驗(yàn)對(duì)象。自第一天訓(xùn)練開(kāi)始分別選擇訓(xùn)練第10天、第17天(消退訓(xùn)練第7天)、第18天(消退后再次聯(lián)合式訓(xùn)練1天)以及對(duì)照組小鼠作為實(shí)驗(yàn)對(duì)象。利用膜片鉗技術(shù)檢測(cè)桶狀皮層和運(yùn)動(dòng)皮層谷氨酸能神經(jīng)元功能的變化。結(jié)果:(1)嗅覺(jué)和胡須刺激聯(lián)合式訓(xùn)練10天后,嗅覺(jué)誘導(dǎo)的胡須擺動(dòng)聯(lián)合式學(xué)習(xí)記憶模型建立;模型建立后給予不配對(duì)刺激消退訓(xùn)練7天后,聯(lián)合式學(xué)習(xí)記憶消退;消退后再次配對(duì)訓(xùn)練1天,聯(lián)合式學(xué)習(xí)記憶重建:與對(duì)照組相比,聯(lián)合式訓(xùn)練10天后實(shí)驗(yàn)組小鼠胡須擺動(dòng)頻率、角度、回縮時(shí)間明顯增加(p0.01);與聯(lián)合式訓(xùn)練第10天相比,消退訓(xùn)練7天后,小鼠胡須擺動(dòng)頻率、角度、回縮時(shí)間明顯減小(p0.01);與消退訓(xùn)練7天后相比,再次給予聯(lián)合式訓(xùn)練1天后,小鼠胡須擺動(dòng)頻率、角度、回縮時(shí)間明顯增加(p0.01)。(2)在聯(lián)合式記憶建立、消退、重建過(guò)程中,桶狀皮層神經(jīng)元功能持續(xù)上調(diào):與對(duì)照組相比,實(shí)驗(yàn)組小鼠聯(lián)合式訓(xùn)練10天后桶狀皮層神經(jīng)元編碼動(dòng)作電位數(shù)量增多(p0.01)、自發(fā)性興奮性突觸后電流(spontaneous excitatory postsynaptic current,s EPSC)發(fā)放頻率和幅度增大(p0.01)、自發(fā)性抑制性突觸后電流(spontaneous inhibitory postsynaptic current,s IPSC)發(fā)放頻率和幅度減小(p0.01)。與聯(lián)合式訓(xùn)練第10天相比,消退訓(xùn)練7天后桶狀皮層在相同的刺激強(qiáng)度下產(chǎn)生的動(dòng)作電位數(shù)量、s EPSC發(fā)放頻率和幅度、s IPSC發(fā)放頻率和幅度都沒(méi)有統(tǒng)計(jì)學(xué)差異;與消退訓(xùn)練7天相比,再次聯(lián)合式訓(xùn)練1天后桶狀皮層神經(jīng)元編碼動(dòng)作電位數(shù)量、s EPSC發(fā)放頻率和幅度、s IPSC發(fā)放頻率和幅度都沒(méi)有統(tǒng)計(jì)學(xué)差異。(3)運(yùn)動(dòng)皮層神經(jīng)元在聯(lián)合式記憶建立時(shí)功能上調(diào),在聯(lián)合式學(xué)習(xí)記憶消退后神經(jīng)元功能下調(diào),在聯(lián)合式學(xué)習(xí)記憶重建后神經(jīng)元功能再次上調(diào):與對(duì)照組相比,實(shí)驗(yàn)組小鼠聯(lián)合式訓(xùn)練10天后運(yùn)動(dòng)皮層神經(jīng)元編碼動(dòng)作電位數(shù)量增多(p0.01),s EPSC發(fā)放頻率和幅度增大(p0.01),自發(fā)性抑制性突觸后電流s IPSC發(fā)放頻率和幅度減小(p0.01);與聯(lián)合式訓(xùn)練第10天相比,消退訓(xùn)練7天后運(yùn)動(dòng)皮層在相同的刺激強(qiáng)度下產(chǎn)生的動(dòng)作電位數(shù)量減少(p0.01)、s EPSC發(fā)放頻率和幅度減小(p0.01)、s IPSC發(fā)放頻率和幅度增加(p0.01);與消退訓(xùn)練7天相比,再次聯(lián)合式訓(xùn)練1天后運(yùn)動(dòng)皮層神經(jīng)元編碼動(dòng)作電位數(shù)量增加(p0.01)、s EPSC發(fā)放頻率和幅度增大(p0.01)、s IPSC發(fā)放頻率和幅度減小(p0.01)。結(jié)論:1.嗅覺(jué)誘導(dǎo)的胡須擺動(dòng)聯(lián)合式學(xué)習(xí)記憶維持與桶狀皮層谷氨酸神經(jīng)元功能上調(diào)有關(guān)。2.嗅覺(jué)誘導(dǎo)的胡須擺動(dòng)聯(lián)合式學(xué)習(xí)記憶消退與運(yùn)動(dòng)皮層谷氨酸能神經(jīng)元功能下調(diào)有關(guān)。
[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.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R338

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