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1、 硕士学位论文 双眼形觉剥夺成年大鼠视皮层可塑性与 CSPGs 和 tPA 表达关系的研究 刘 瑶 刘 瑶 指导教师 阴正勤 教授 第三军医大学西南医院眼科，400038 申请学位级别 硕士科学学位 专业名称眼科学 论文提交日期 2011 年 11 月 论文答辩日期2011 年 11 月 学位授予单位和日期 第三军医大学 2011 年 月 答 辩 委 员 会 主 席 评 阅 人 二一一年十一月 分类号分类号 密密 级级 公公 开开 学学 号号 2008151 目 录 主要英文缩写词表 1 英文摘要 2 中文摘要 5 论文正文 双眼形觉剥夺成年大鼠视皮层可塑性与双眼形觉剥夺成年大鼠视皮层可塑性与
2、 CSPGs 和和 tPA 表达关系的研究表达关系的研究 . 8 前 言 8 第一部分第一部分 建立双眼形觉剥夺成年大鼠视皮层可塑性“再激活”模型建立双眼形觉剥夺成年大鼠视皮层可塑性“再激活”模型 11 材料和方法.11 结 果.14 讨 论.21 小 结.23 第二部分第二部分 双眼形觉剥夺成年大鼠视皮层双眼形觉剥夺成年大鼠视皮层 tPA 表达及活性变化的研究表达及活性变化的研究24 材料和方法.24 结 果.28 讨 论.33 小 结.34 第三部分第三部分 双眼形觉剥夺成年大鼠视皮层双眼形觉剥夺成年大鼠视皮层 tPA 与与 CSPGs 表达相互关系的研究表达相互关系的研究35 材料和方法
3、.35 结 果.37 讨 论.43 小 结.45 全文结论.46 致 谢.47 参考文献.48 文献综述 视皮层内的抑制性环路对视皮层可塑性的起始和终止的影响视皮层内的抑制性环路对视皮层可塑性的起始和终止的影响.53 参考文献.56 在读期间发表的文章 .59 第三军医大学硕士学位论文 1 主要英文缩写词表 缩写 英文名称 中文名称或注释 BFD binocular form deprivation 双眼形觉剥夺 CSPGs chondroitin sulphate proteoglycans 硫酸软骨素蛋白多糖 ECM extracellular matrix 细胞外基质 FVEP flas
4、h-visual evoked potential 闪光视觉诱发电位 GABA -amino butyric acid -氨基丁酸 MD monocular deprivation 单眼剥夺 NMDA N-methyl-D-aspartate N-甲基-D-天冬氨酸 PBS Phosphate buffered saline 磷酸盐缓冲液 PFA paraformaldehyed 多聚甲醛 PNNs perineuronal net 神经元周围网络 PVEP pattern visual evoked potential 图形视觉诱发电位 PW postnatal weeks 出生后周龄 tP
5、A tissue-type plasminogen activator 组织型纤溶酶原激活剂 VEPs visual evoked potentials 视觉诱发电位 第三军医大学硕士学位论文 2 The visual cortex plasticity and relationship between CSPGs and tPA in adult visual cortex after Binocular form deprivation Abstract After human and the mammalians born, visual system regulated the con
6、nection of neurons and structures of synaptics according to the visual environment, which is called the critical period of visual cortex plasticity. Researches in the past showed that amblyopia can be treated only in this critical period. Present researches showed that visual plasticity in the adult
7、 is just inhibited but not disappeared and it can be “reactivated” in some specific conditions. In 2002, Pizzorusso degraded the CSPGs in adult rats visual cortex by using chABC enzymes, which first successfully recovered adult rats visual cortex plasticity, and raised the amblyopic eyes visual acui
8、ty by deprivating inputs of the fellow eye. This study first confirmed that the plasticity of adult visual cortex can be “reactivated”. Thereafter, some studies which used 10 days dark rearing、 environment enrichment、 the antidepressant drugs and reducing intracortical inhibition in adult rats can a
9、lso recover adult visual cortex plasticity. Our researches in the past found that binocular form deprivation can inhibite the development of GABA circuit, reduce the strength of neuron synapses transmission, and regulate the expression and distribution of inhibiting and exciting neurotransmitter rec
10、eptors in adult visual cortex. Researches have showed that maturing of PNNs constituted by CSPGs can promote the maturing of GABA inhibition circuit, but degradation of PNNs can reduce the inhibition of GABA circuit. Therefore, reducing of PNNs is one of the ways to turn down inhibition of GABA circ
11、uit and whether it can also be caused by binocular form deprivation(BFD) is not known. TPA is one of the natural factors in CSPGs degradation, and plays an important role in mediating the ocular dominance shift in response to monocular deprivation. Researches have showed that, the activity of tPA 第三
12、军医大学硕士学位论文 3 increased in contralateral visual cortex caused by monocular deprivation in critical period. Whether it can also happen in BFD is not known. Overall, we made this hypothesis: tPA activity increases in visual cortex in response to binocular form deprivation in adult rats, which degrades
13、CSPGs, reduses expression of PNNs, and then “reactivates” adult visual cortex plasticity. Therefore we applied the following methods to investigate this hypothesis: 1、Pattern visual evoked potentials(PVEPs) was applied to detect the OD shift in order to find out the end time of visual cortex plastic
14、ity and the proper time to reactive adult visual cortex plasticity by binocular form deprivation in adult rats. Results：(1) Brief 3 days monocular deprivation caused the OD shift in PW3、PW4 and PW5 rats, which showed the visual cortex plasticity existed. (2) The OD shift could not be observed in rat
15、s after PW6 after brief 3 days monocular deprevation, which showed the visual cortex plasticity was inhibited. (3) A significant OD shift was observed in PW7 rats after 14 days BFD in response to brief 3 days monocular deprivation, which demonstrated a steady reactivation of visual cortex plasticity
16、 in adult rats. (4) 14 days BFD was taken as the model in the following experiments. 2、 With the use of immunofluorescence histochemistry, immunoblot and ELISA kit, we detected tPA expression and activity in visual cortex during the critical period and 14 days BFD in adult rats. Results：(1) There were only a few cells expressing tPA in visual cortex in PW1, and increased significantly after eyes open. The expression of tPA depended on the visual experience. (
