發(fā)布時(shí)間：2018-10-18 15:40

【摘要】：目的：丘腦前核（anterior thalamic nuclei，ATN）是位于間腦背側(cè)丘腦前部的重要聯(lián)絡(luò)性核團(tuán)，是Papez回路的重要組成部分，由三個(gè)亞核組成，分別為丘腦前核背側(cè)核（AD），丘腦前核腹側(cè)核（AV）及丘腦前核內(nèi)側(cè)核(AM）。已有實(shí)驗(yàn)證明丘腦前核背側(cè)核及丘腦前核腹側(cè)核在學(xué)習(xí)記憶過(guò)程中同樣發(fā)揮積極作用，但對(duì)于其參與學(xué)習(xí)記憶機(jī)制的研究還很少。本研究旨在通過(guò)應(yīng)用免疫組化技術(shù)觀察Morris水迷宮訓(xùn)練建模成功大鼠的丘腦前核蛋白激酶A催化亞基β（PKA-Cβ）的表達(dá)探討其在丘腦前核學(xué)習(xí)記憶中的可能作用，進(jìn)一步揭示丘腦前核在學(xué)習(xí)記憶中從發(fā)揮作用的相關(guān)分子機(jī)制。 方法：本實(shí)驗(yàn)采用30只成年雄性Wistar大鼠，體重200-250g，隨即分配至實(shí)驗(yàn)組與對(duì)照組。實(shí)驗(yàn)組經(jīng)5天Morris水迷宮實(shí)驗(yàn)建模形成空間記憶，對(duì)照組單純給予相同條件的環(huán)境刺激但不給予學(xué)習(xí)記憶訓(xùn)練。于第5天空間探索試驗(yàn)后兩小時(shí)內(nèi)行4%多聚甲醛心臟灌流，固定，取腦，4°C過(guò)夜。次日用免疫組織化學(xué)染色(immunohistochemistry, IHC)方法觀察大鼠丘腦前核內(nèi)蛋白激酶A催化亞基β的表達(dá)及分布情況，顯微拍照并行圖片分析。Morris水迷宮行為學(xué)實(shí)驗(yàn)數(shù)據(jù)及照片分析數(shù)據(jù)均使用SPSS軟件進(jìn)行單因素方差分析，P＜0.05為差異有統(tǒng)計(jì)學(xué)意義。 結(jié)果：Morris水迷宮行為學(xué)實(shí)驗(yàn)中，通過(guò)對(duì)實(shí)驗(yàn)組大鼠第三天和第五天的獲得性訓(xùn)練試驗(yàn)及第六天的空間探索實(shí)驗(yàn)的逃避潛伏期行數(shù)據(jù)分析，發(fā)現(xiàn)第五天與第三天的逃避潛伏期相比明顯縮短（P0.05），第五天與第六天的逃避潛伏期間沒(méi)有顯著差異。免疫組化染色中，光鏡下可見(jiàn)蛋白激酶A催化亞基β在實(shí)驗(yàn)組大鼠丘腦前核內(nèi)的細(xì)胞核及神經(jīng)纖維中較清晰的表達(dá)。通過(guò)對(duì)陽(yáng)性表達(dá)產(chǎn)物的平均光密度值分析，實(shí)驗(yàn)組表達(dá)強(qiáng)于對(duì)照組，，有顯著差異（P0.05）；實(shí)驗(yàn)組AD區(qū)表達(dá)弱于AV區(qū)，有顯著差異（P0.05）；對(duì)照組AD區(qū)與AV區(qū)差異不顯著（P0.05）。 結(jié)論：在學(xué)習(xí)記憶過(guò)程中大鼠丘腦前核內(nèi)蛋白激酶A催化亞基β的表達(dá)增加，提示丘腦前核參與空間學(xué)習(xí)記憶機(jī)制與蛋白激酶A催化亞基β相關(guān)。
[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 擄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.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R338

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