本文選題：桶狀皮層 + GABA能神經(jīng)元��； 參考：《蚌埠醫(yī)學院》2012年碩士論文

【摘要】：目的：建立小鼠觸須感覺信息輸入減少的動物模型，分析觸須感覺信息輸入減少后，桶狀皮層（Barrel Cortex）錐體神經(jīng)元和GABA能神經(jīng)元內(nèi)在特性和神經(jīng)元群集發(fā)放動作電位編碼功能的變化，探討神經(jīng)網(wǎng)絡功能的可塑性機制。 方法：1.小鼠觸須感覺信息輸入減少模型的建立：將FVB-Tg（Gad GFP）4570Swn/J小鼠右側(cè)嘴角部全部觸須修剪至小于1mm，每24小時修剪一次，從出生后7天開始修剪持續(xù)到出生后17天。 2.在上述處理10天以后，應用T迷宮實驗檢測小鼠嗅覺功能變化，實驗中小鼠移動到食物一側(cè)的成功率作為鑒別其嗅覺功能上調(diào)的指標。 3.膜片鉗電生理實驗：取出生1周后小鼠，連續(xù)造模后10天，用異氟烷吸入麻醉，迅速將大腦分離，做冠狀切片（400μm），然后將腦片置于氧合的人工腦脊液(ASCF)中，25C下孵育1-2小時后轉(zhuǎn)移至灌流槽，在31C下對腦片進行灌流；采用IR-DIC光學顯微鏡(Nikon E600FN)熒光顯微鏡定位大腦桶狀皮層綠色熒光蛋白標記的GABA能神經(jīng)元和錐體神經(jīng)元；運用Axoclamp-200B放大器，電流鉗模式全細胞記錄方法記錄大腦桶狀皮層GABA能神經(jīng)元和錐體神經(jīng)元的動作電位，測量其發(fā)放容量及閾電位(threshold potentials, Vts)，電壓鉗模式全細胞記錄方法采集GABA能神經(jīng)元sIPSCs（自發(fā)性抑制性突觸后電流）包括電流間距（Interval of Events，IE）和電流幅度（Amplitude）數(shù)據(jù)，信號數(shù)據(jù)輸入Clampex10.1進行處理分析。結(jié)果：1.造模組小鼠對食物的正確選擇成功率明顯高于正常組小鼠；2.應用膜片鉗方法，，電流鉗模式實驗結(jié)果發(fā)現(xiàn)，與正常組相比，造模組桶狀皮層錐體神經(jīng)元群集發(fā)放動作電位的發(fā)放容量顯著增加，動作電位間距ISI縮短(P0.05)，閾電位(Vts)降低(P0.05)；GABA能神經(jīng)元群集發(fā)放動作電位的發(fā)放容量顯著降低，動作電位間距ISI延長(P0.05)，閾電位(Vts)升高(P0.05)；3.電壓鉗模式實驗結(jié)果發(fā)現(xiàn)，與正常組相比，造模組桶狀皮層GABA能神經(jīng)元的sIPSCs的電流間距延長（P0.05），電流幅度降低(P0.05)。 結(jié)論：1.觸須感覺信息輸入減少后，小鼠嗅覺功能上調(diào)；2.小鼠觸須感覺信息輸入減少，桶狀皮層錐體神經(jīng)元功能上調(diào)，GABA能神經(jīng)元功能下調(diào)；桶狀皮層興奮性神經(jīng)元功能上調(diào)和抑制性神經(jīng)元功能下調(diào)導致神經(jīng)網(wǎng)絡整體功能增強。
[Abstract]:Objective: to establish an animal model of decreased sensory information input in mouse tentacles, and to analyze the reduction of sensory information input in mouse tentacles. The intrinsic characteristics of pyramidal neurons and GABA neurons in barrel-Cortex-pyramidal neurons and the changes in the coding function of the action potential of neuron clusters were studied to explore the plasticity mechanism of neural network function. Method 1: 1. Establishment of a model for reducing the input of sensory information in mouse tentacles: all the tentacles in the right mouth corner of FVB-Tg(Gad GFP)4570Swn/J mice were trimmed to less than 1 mm, trimmed every 24 hours, from 7 days after birth to 17 days after birth. 2. After 10 days of treatment, T maze test was used to detect the changes of olfactory function in mice. The success rate of mice moving to one side of food was used as an index to identify the up-regulation of olfactory function. 3. Patch clamp electrophysiological experiment: the mice were taken out one week after birth. After 10 days of continuous modeling, isoflurane was inhaled to anesthetize the brain, and the brain was separated quickly. The brain slices were incubated at 25C for 1-2 hours and then transferred to the perfusion trough. The slices were perfused at 31C, then the brain slices were incubated in the oxygenated artificial cerebrospinal fluid (ASCF) for 1 to 2 hours. The IR-DIC optical microscope and Nikon E600FN fluorescence microscope were used to locate the GABA neurons and pyramidal neurons labeled with green fluorescent protein (GFP) in the barrelled cortex, and the Axoclamp-200B amplifier was used. The action potentials of GABA neurons and pyramidal neurons in the barrelled cortex were recorded by current clamp mode whole-cell recording method. The release capacity, threshold potential, voltage clamp mode and voltage clamp mode whole-cell recording were used to collect the data of sIPSCs (spontaneous inhibitory postsynaptic currents) including current spacing interval of events IEE and amplitude of current Amplitudee. The signal data is input into Clampex10.1 for processing and analysis. The result is 1: 1. The success rate of food selection in model group was significantly higher than that in normal group. By using patch clamp method and current clamp mode experiment, it was found that compared with the normal group, the discharge capacity of the pyramidal pyramidal neurons in the bucket cortex in the model group was significantly increased. Action potential spacing (ISI) shortened P0.05N and threshold potential (VtsS) decreased significantly the release capacity of P0.05GABA-neuron cluster action potential (P0.05N). The interval of action potential (ISI) was prolonged (P0.05N), and the threshold potential (Vts) increased (P0.05) ~ (3). The results of voltage clamp mode experiment showed that compared with the normal group, the sIPSCs current spacing of the barrelled cortical GABA neurons in the model group was prolonged (P0.05) and the current amplitude was decreased (P0.05). Conclusion 1. The olfactory function of mice was upregulated by tactile sensory information input. The sensory information input of mouse tentacles decreased, the function of pyramidal neurons in the bucket cortex increased and the function of GABA neurons down-regulated, while the function of excitatory neurons in the barrelled cortex and the down-regulation of the inhibitory neurons resulted in the enhancement of the overall function of the neural network.
【學位授予單位】：蚌埠醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R338.1

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