大鼠丘脑前核内蛋白激酶A催化亚基β在学习记忆中的表达
发布时间：2018-10-18 15:40
【摘要】：目的：丘脑前核（anterior thalamic nuclei，ATN）是位于间脑背侧丘脑前部的重要联络性核团，是Papez回路的重要组成部分，由三个亚核组成，分别为丘脑前核背侧核（AD），丘脑前核腹侧核（AV）及丘脑前核内侧核(AM）。已有实验证明丘脑前核背侧核及丘脑前核腹侧核在学习记忆过程中同样发挥积极作用，但对于其参与学习记忆机制的研究还很少。本研究旨在通过应用免疫组化技术观察Morris水迷宫训练建模成功大鼠的丘脑前核蛋白激酶A催化亚基β（PKA-Cβ）的表达探讨其在丘脑前核学习记忆中的可能作用，进一步揭示丘脑前核在学习记忆中从发挥作用的相关分子机制。 方法：本实验采用30只成年雄性Wistar大鼠，体重200-250g，随即分配至实验组与对照组。实验组经5天Morris水迷宫实验建模形成空间记忆，对照组单纯给予相同条件的环境刺激但不给予学习记忆训练。于第5天空间探索试验后两小时内行4%多聚甲醛心脏灌流，固定，取脑，4°C过夜。次日用免疫组织化学染色(immunohistochemistry, IHC)方法观察大鼠丘脑前核内蛋白激酶A催化亚基β的表达及分布情况，显微拍照并行图片分析。Morris水迷宫行为学实验数据及照片分析数据均使用SPSS软件进行单因素方差分析，P＜0.05为差异有统计学意义。 结果：Morris水迷宫行为学实验中，通过对实验组大鼠第三天和第五天的获得性训练试验及第六天的空间探索实验的逃避潜伏期行数据分析，发现第五天与第三天的逃避潜伏期相比明显缩短（P0.05），第五天与第六天的逃避潜伏期间没有显著差异。免疫组化染色中，光镜下可见蛋白激酶A催化亚基β在实验组大鼠丘脑前核内的细胞核及神经纤维中较清晰的表达。通过对阳性表达产物的平均光密度值分析，实验组表达强于对照组，，有显著差异（P0.05）；实验组AD区表达弱于AV区，有显著差异（P0.05）；对照组AD区与AV区差异不显著（P0.05）。 结论：在学习记忆过程中大鼠丘脑前核内蛋白激酶A催化亚基β的表达增加，提示丘脑前核参与空间学习记忆机制与蛋白激酶A催化亚基β相关。
[Abstract]:Objective: the anterior thalamic nucleus (anterior thalamic nuclei,ATN) is an important conjunctive nucleus located in the anterior part of the dorsal diencephalon. It is an important component of the Papez loop and consists of three subnuclei. (AD), (AV) of ventral nucleus of anterior nucleus of thalamus and (AM). Of medial nucleus of anterior nucleus of thalamus It has been proved that the dorsal nucleus of the anterior thalamic nucleus and the ventral nucleus of the anterior nucleus of the thalamus also play an active role in the process of learning and memory, but little research has been done on their involvement in the mechanism of learning and memory. The purpose of this study was to observe the expression of prokaryotic protein kinase A catalytic subunit 尾 (PKA-C 尾) in the thalamus of Morris water maze training rats by using immunohistochemical technique to explore its possible role in the learning and memory of the anterior thalamus. Furthermore, the molecular mechanism of the role of the anterior thalamic nucleus in learning and memory is revealed. Methods: 30 adult male Wistar rats, weighing 200-250 g, were assigned to the experimental group and the control group. After 5 days of Morris water maze experiment, the experimental group was modeled to form spatial memory, while the control group was given only environmental stimulation under the same conditions but not learning and memory training. The heart was perfused with 4% paraformaldehyde in 2 hours after space exploration test on the 5th day, fixed, brain was taken, and 4 掳C was used overnight. The expression and distribution of protein kinase A catalytic subunit 尾 in rat anterior thalamus were observed by immunohistochemical staining (immunohistochemistry, IHC) method the next day. The data of Morris water maze behavior experiment and photo analysis were analyzed by SPSS software, and the difference was statistically significant (P < 0. 05). Results: in the behavior experiment of Morris water maze, the escape latency of the third and fifth day of the experimental group and the sixth day of space exploration experiment were analyzed. It was found that the escape latency of the fifth day was significantly shorter than that of the third day (P0.05), but there was no significant difference between the fifth day and the sixth day. The expression of protein kinase A catalytic subunit 尾 in the nucleus and nerve fibers of the anterior thalamic nucleus of the experimental group was clear under light microscope. Through the analysis of the average optical density of the positive expression products, the expression of AD in the experimental group was stronger than that in the control group (P0.05); the expression of AD in the experimental group was weaker than that in the AV region (P0.05); in the control group, there was no significant difference between the AD region and the AV region (P0.05). Conclusion: during learning and memory, the expression of protein kinase A catalytic subunit 尾 in the anterior nucleus of the thalamus is increased, suggesting that the anterior thalamus nucleus is involved in the spatial learning and memory mechanism and the protein kinase A catalytic subunit 尾 is involved in the process of learning and memory.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R338
本文编号：2279579
[Abstract]:Objective: the anterior thalamic nucleus (anterior thalamic nuclei,ATN) is an important conjunctive nucleus located in the anterior part of the dorsal diencephalon. It is an important component of the Papez loop and consists of three subnuclei. (AD), (AV) of ventral nucleus of anterior nucleus of thalamus and (AM). Of medial nucleus of anterior nucleus of thalamus It has been proved that the dorsal nucleus of the anterior thalamic nucleus and the ventral nucleus of the anterior nucleus of the thalamus also play an active role in the process of learning and memory, but little research has been done on their involvement in the mechanism of learning and memory. The purpose of this study was to observe the expression of prokaryotic protein kinase A catalytic subunit 尾 (PKA-C 尾) in the thalamus of Morris water maze training rats by using immunohistochemical technique to explore its possible role in the learning and memory of the anterior thalamus. Furthermore, the molecular mechanism of the role of the anterior thalamic nucleus in learning and memory is revealed. Methods: 30 adult male Wistar rats, weighing 200-250 g, were assigned to the experimental group and the control group. After 5 days of Morris water maze experiment, the experimental group was modeled to form spatial memory, while the control group was given only environmental stimulation under the same conditions but not learning and memory training. The heart was perfused with 4% paraformaldehyde in 2 hours after space exploration test on the 5th day, fixed, brain was taken, and 4 掳C was used overnight. The expression and distribution of protein kinase A catalytic subunit 尾 in rat anterior thalamus were observed by immunohistochemical staining (immunohistochemistry, IHC) method the next day. The data of Morris water maze behavior experiment and photo analysis were analyzed by SPSS software, and the difference was statistically significant (P < 0. 05). Results: in the behavior experiment of Morris water maze, the escape latency of the third and fifth day of the experimental group and the sixth day of space exploration experiment were analyzed. It was found that the escape latency of the fifth day was significantly shorter than that of the third day (P0.05), but there was no significant difference between the fifth day and the sixth day. The expression of protein kinase A catalytic subunit 尾 in the nucleus and nerve fibers of the anterior thalamic nucleus of the experimental group was clear under light microscope. Through the analysis of the average optical density of the positive expression products, the expression of AD in the experimental group was stronger than that in the control group (P0.05); the expression of AD in the experimental group was weaker than that in the AV region (P0.05); in the control group, there was no significant difference between the AD region and the AV region (P0.05). Conclusion: during learning and memory, the expression of protein kinase A catalytic subunit 尾 in the anterior nucleus of the thalamus is increased, suggesting that the anterior thalamus nucleus is involved in the spatial learning and memory mechanism and the protein kinase A catalytic subunit 尾 is involved in the process of learning and memory.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R338
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