精神分裂症小鼠脑内GABA能神经元的变化

发布时间：2018-06-08 19:30

  本文选题：精神分裂症 + 前额皮质　； 参考：《宁夏医科大学》2012年硕士论文

【摘要】：目的观察小鼠在发生精神分裂症症状后，刻板行为及共济失调行为的变化情况，前额皮质（PFC）和海马（HIP）CA1、CA2、CA3内GABA的变化情况。 方法应用C57BL/6及GAD67-GFP基因敲入小鼠连续两周腹腔注射不同剂量的地卓西平马来酸盐(MK801)（0.3mg/kg,0.6mg/kg,1mg/kg），进行精神分裂症动物模型的制备。对注药后3、5、7、10、14天小鼠的行为进行录像，对小鼠的共济失调、刻板运动行为等进行分析、评估；小鼠的精神分裂症模型成功后，采用免疫荧光染色方法，观察PFC和HIP的CA1、CA2、CA3内GABA能神经元以及GABA能神经元亚群标志物CB、CR、PV的变化；用RT-PCR方法，观察上述脑区GAD67mRNA表达水平的改变。 结果（1）行为学改变。○1刻板运动评估将0.3mg/kg、0.6mg/kg、1mg/kg三个剂量组小鼠的刻板运动评分进行比较，结果显示，0.6mg/kg、1mg/kg剂量组小鼠的刻板运动评分均较0.3mg/kg剂量组的显著增高，分别增高37.9%、38.1%，而0.6mg/kg剂量组与1mg/kg剂量组之间评分无明显差异；对给药后第3、5、7、10、14天刻板运动进行评分，随着给药天数的增加，各剂量组小鼠刻板运动评分均在逐渐减低。○2共济失调评估将0.3mg/kg、0.6mg/kg、1mg/kg三个剂量组小鼠的共济失调评分进行比较，结果显示，1mg/kg剂量组小鼠的刻板运动评分显著高于0.3mg/kg剂量组的评分，增高36.4%，而0.3mg/kg剂量组与0.6mg/kg剂量组之间，1mg/kg剂量组与0.6mg/kg剂量组之间评分无明显差异；对给药后第3、5、7、10、14天共济失调进行评分，随着给药天数的增加，各剂量组小鼠共济失调评分均在逐渐减低。（2）GABA能神经元的变化。形态学方面。○1PFC中，模型组和对照组小鼠各层均可见GFP阳性细胞，且在Ⅱ-Ⅲ层分布较多，但单位面积阳性细胞数与对照组无明显差异；PV阳性细胞分布于Ⅲ-Ⅴ层，CB和CR阳性细胞主要分布于Ⅱ-Ⅲ层，但单位面积内小清蛋白（PV）、钙结合蛋白（CB）、钙视网膜蛋白（CR）阳性细胞数目与对照组小鼠相比无明显变化。○2HIP的CA1、CA2、CA3区，GFP阳性细胞在各层均有分布，但单位面积内阳性细胞数与对照组相比无明显变化；PV、CB阳性细胞主要分布于CA1的起始层，单位面积内阳性细胞数与对照组无明显差异；CR阳性细胞在CA1区较对照组明显增多，，而在CA2及CA3区单位面积内阳性细胞数较对照组无明显差异。RT-PCR结果。精神分裂症模型组小鼠，PFC中，GAD67mRNA的表达量较盐水对照组明显减少，而在海马组织中，GAD67mRNA的表达量与盐水对照组无明显差异。 结论小鼠注射MK801后刻板运动及共济失调行为具有剂量依赖性；长期给予1mg/kg及其以内剂量的MK801可能使小鼠的刻板运动及共济失调运动行为减弱，其可能与小鼠体内递质的自我调节及小鼠的行为适应有关；精神分裂症时在PFC及HIP的CA1、CA2、CA3脑区抑制性神经递质GABA在蛋白水平未见明显改变；在mRNA水平上，GABA在PFC减少。这种变化与精神分裂症的关系尚有待进一步的研究。
[Abstract]:Objective to observe the changes of stereotypical behavior and ataxia behavior in mice after the onset of schizophrenia. Methods using C57BL / 6 and GAD67-GFP knockout mice for two consecutive weeks intraperitoneally injected different doses of didroxepine maleate (MK801) 0.3 mg / kg 0.6 mg / kg 路kg ~ (-1) mg 路kg ~ (-1) 路L ~ (-1) mg 路kg ~ (-1) ~ (-1) mg 路kg ~ (-1)? The behavior of mice was recorded on 3 days after injection, and the ataxia and stereotypical motor behavior of mice were analyzed and evaluated. After the success of the model of schizophrenia in mice, immunofluorescence staining was used. To observe the changes of GABA neurons and the subsets of GABA neurons in CA1, CA2 and CA3 of PFC and hip, and to observe the changes of CBB CRN PV, a subgroup marker of GABA neurons in PFC and hip. The changes of GAD67 mRNA expression in the above brain region were observed. Results 1) Behavioral changes. 01 stereotype motor evaluation. The stereotype motor scores of 0.3 mg / kg ~ 0.6 mg 路kg ~ (-1) 路kg ~ (-1) mg 路kg ~ (-1) were compared. The results showed that the scores of stereotype motor of mice in 0.6 mg / kg / kg group were significantly higher than those in 0.3mg/kg group, which were 37.9% and 38.1%, respectively, but there was no significant difference between the 0.6mg/kg group and the 1mg/kg group. With the increase of administration days, the stereotypical motor score of mice in each dose group was gradually decreased .02. The ataxia score of 0.3 mg / kg + 0.6 mg / kg + 1 mg / kg group was compared with that of the three dose groups. The results showed that the stereotype motor score of mice in the 1 mg / kg group was significantly higher than that in the 0.3mg/kg group and increased 36.4%, but there was no significant difference between the 1 mg / kg group and the 0.6mg/kg group in the 0.3mg/kg dose group and the 0.6mg/kg dose group. The ataxia scores of mice at day 3, 5, 7, 10 and 14 days after administration were evaluated. With the increase of administration days, the ataxia scores of mice in each dose group were gradually decreased. The changes of GABA neurons in each dose group were gradually decreased. In morphological aspect, GFP positive cells were found in each layer of model group and control group, and there were more GFP positive cells in layer 鈪

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