条件恐惧刺激对成年大鼠杏仁核干细胞增殖及分化为神经元的影响

发布时间：2018-02-13 18:52

  本文关键词： 条件恐惧刺激 杏仁核 BrdU NeuN 免疫组化　出处：《中南大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的：检测条件恐惧刺激后大鼠杏仁核区神经干细胞的数量及神经干细胞分化为神经元的量的变化,探讨条件恐惧刺激后大鼠杏仁核区神经干细胞的增殖及分化与恐惧记忆的关系。 方法：1.应用足底电击配对单调声刺激的方法建立大鼠条件恐惧记忆模型,并以呆立时间(freezing time)评价大鼠对条件恐惧刺激事件的记忆。2.应用免疫组化方法检测条件恐惧刺激后不同时间点(1、7、14、21、30、60天)大鼠杏仁核区BrdU(外源性神经干细胞标记物)阳性细胞的表达量。3.应用免疫荧光双标记法检测条件恐惧刺激后不同时间点大鼠杏仁核区BrdU与NeuN(神经元标记物)双标阳性细胞的量。4.应用SPSS15.0统计软件进行统计学分析。 结果：1.行为学检测结果显示：1、7、14、21、30、60天时间点的实验组大鼠在被重新放入电击箱给予单调声刺激后,八分钟内的呆立时间明显长于相应时间点的对照组,差异具有显著性(P0.01)Do。2. BrdU的免疫组织化学结果显示：在大鼠杏仁核区可见典型的BrdU阳性细胞,计数杏仁核区BrdU阳性细胞,发现条件恐惧刺激后各时间点实验组大鼠杏仁核区BrdU免疫阳性细胞数少于相应对照组,其中1、7、14、21、30天差异具有显著性(P0.05),60天实验组与对照组的差异没有统计学意义。3.BrdU与NeuN的免疫荧光双标结果显示：在大鼠杏仁核区可见明显的BrdU与NeuN双标阳性细胞,计数杏仁核区BrdU阳性细胞及BrdU与NeuN的双标阳性细胞,发现条件恐惧刺激后各时间点实验组大鼠杏仁核区BrdU和NeuN免疫荧光双标阳性细胞数与BrdU阳性细胞总数的比值小于相应对照组,差异具有显著性(P0.05)。 结论：条件恐惧刺激后大鼠呆立时间明显延长,且杏仁核区细胞增殖减少。以上结果提示条件恐惧刺激有可能影响大鼠杏仁核区的神经元新生。
[Abstract]:Objective: to detect the number of neural stem cells (NSCs) in amygdala and the number of neural stem cells (NSCs) differentiating into neurons after conditioned fear stimulation. To investigate the relationship between fear memory and proliferation and differentiation of neural stem cells in amygdala region after conditioned fear stimulation. Methods 1. The conditioned fear memory model of rats was established by plantar shock matching monotone sound stimulation. The memory of conditioned fear stimulation events in rats was evaluated by standing time and freezing time. 2. Immunohistochemical method was used to detect BrdU (exogenous neural stem cell marker) positive in amygdaloid nucleus of rats at different time points after conditioned fear stimulation. The expression of sex cells was measured by immunofluorescence double labeling method. The amount of BrdU and Neun (neuron marker) double labeled positive cells in amygdaloid nucleus of rats at different time points after conditioned fear stimulation was detected. 4. SPSS15.0 statistical software was used for statistical analysis. Results: 1.Behavioral tests showed that the rats in the experimental group at 30 days after being put back into the electric shock box for monotone sound stimulation were significantly longer in eight minutes than those in the control group at the corresponding time point. The immunohistochemical results of BrdU showed that typical BrdU positive cells were found in the amygdala of rats, and the BrdU positive cells in the amygdala were counted. It was found that the number of BrdU immunoreactive cells in the amygdala of the experimental group was less than that of the corresponding control group at each time point after conditioned fear stimulation. There was no significant difference between the experimental group and the control group on the 30th day. 3. The immunofluorescence double labeling results of BrdU and NeuN showed that there were obvious double labeled BrdU and NeuN positive cells in the amygdala of rats. BrdU positive cells and double labeled positive cells of BrdU and NeuN in amygdala were counted. It was found that the ratio of the number of BrdU and NeuN double labeled positive cells to the total number of BrdU positive cells in the amygdaloid nucleus of the experimental group was lower than that of the corresponding control group at each time point after conditioned fear stimulation, and the difference was significant (P 0.05). Conclusion: the experimental results suggest that conditioned fear stimulation may affect the neuronal growth of amygdaloid area in rats.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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