小鼠慢性應(yīng)激模型海馬中tau蛋白磷酸化及其對線粒體軸漿運輸?shù)挠绊?/H1>
發(fā)布時間：2018-04-13 02:10
本文選題：抑郁(Depression) + 母子剝奪(Maternal ； 參考：《中國科學技術(shù)大學》2011年博士論文
【摘要】：Tau蛋白是一種微管相關(guān)蛋白,其磷酸化受到嚴格調(diào)控,在神經(jīng)元中和軸漿運輸密切相關(guān)。據(jù)研究報道,急性應(yīng)激中,嚙齒類動物海馬中tau蛋白的磷酸化和下丘腦-垂體-腎上腺(Hypothalamo-Pituitary-Adrenal,HPA)軸的過度激活密切相關(guān)。而在抑郁病人中,HPA軸的過度激活是基本的特征之一。因此,在抑郁癥中,tau蛋白的磷酸化情況以及和軸漿運輸?shù)年P(guān)系需要進一步研究。我們的假設(shè)是:在抑郁模型中,HPA軸的過度激活會引起tau蛋白的過度磷酸化并進一步影響到軸漿運輸。 為驗證該假設(shè),我們首先建立了母子剝奪抑郁模型,對其進行了行為學測試,并檢測了HPA軸活性。然后檢測了海馬中tau蛋白的磷酸化水平。為了研究軸漿運輸?shù)淖兓?我們選擇了檢測海馬突觸體中線粒體水平的變化。結(jié)合CRH(Corticotropin-Releasing Hormone,CRH)受體1(CRH Receptor1,CRHR1)和糖皮質(zhì)激素受體(Glucocorticoid Receptor,GR)的拮抗劑,研究了CRH直接的中樞作用機制和HPA軸的反饋機制在tau蛋白磷酸化和線粒體運輸中的作用,。為了進一步檢測tau蛋白磷酸化和線粒體運輸?shù)年P(guān)系,我們用實時成像技術(shù),觀察當原代神經(jīng)元中tau蛋白磷酸化水平發(fā)生變化時線粒體運輸?shù)膭討B(tài)變化。結(jié)果發(fā)現(xiàn): 1.母子剝奪抑郁模型具有典型的抑郁樣行為和過度激活的HPA軸。 在曠場、糖水偏好和明暗箱行為學測試中,抑郁模型具有典型的抑郁樣行為。和對照組相比,抑郁模型具有顯著升高的血漿cortitosterone和下丘腦CRH mRNA水平。 2.抑郁模型海馬中tau蛋白的磷酸化水平顯著升高。 和對照組相比,抑郁鼠的海馬中tau蛋白的總表達水平?jīng)]有顯著變化,但在在AT-8磷酸化位點顯著升高,在tau-1非磷酸化位點顯著降低。 3.抑郁模型海馬突觸體中的線粒體水平顯著升高。 和對照組相比,抑郁模型海馬突觸體中的線粒體水平顯著升高。 4. CRHR1而不是GR介導了tau蛋白的磷酸化水平升高和突觸體中線粒體水平的升高。 CRHR1的拮抗劑CP154526能顯著降低tau蛋白的磷酸化,并能恢復突觸體中線粒體的水平,而GR的拮抗劑RU486卻沒有這些作用。 5.在原代神經(jīng)元中,當tau蛋白磷酸化水平降低時,線粒體的軸漿運輸水平下調(diào)。我們用鋰鹽抑制Gsk3beta的活性從而降低原代神經(jīng)元中tau蛋白的磷酸化水 平,我們觀察到線粒體的軸漿運輸水平顯著下降。 上述結(jié)果表明,在抑郁模型中CRHR1介導了tau蛋白的過度磷酸化,同時伴隨著突觸體中線粒體水平的升高。這向我們揭示,在抑郁模型中,CRH的中樞作用參與了線粒體運輸?shù)恼{(diào)控,這可能為研究抑郁發(fā)病機制提出了一條新思路。
[Abstract]:Tau protein is a microtubule-associated protein, its phosphorylation is strictly regulated, transport in neurons and axonal transport is closely related.It has been reported that the phosphorylation of tau protein in the hippocampus of rodents is closely related to the hyperactivation of the hypothalamo-Pituitary-Adrenalar axis during acute stress.Hyperactivation of HPA axis is one of the basic characteristics in depressive patients.Therefore, the phosphorylation of tau and its relationship with axonal transport in depression need further study.Our hypothesis is that excessive activation of the HPA-axis in depression models leads to excessive phosphorylation of tau proteins and further affects axonal transport.To verify this hypothesis, we first established a model of mother-son deprivation depression, tested it by behavioral tests, and measured the activity of the HPA axis.Then the phosphorylation level of tau protein in hippocampus was detected.In order to study the changes of axonal transport, we chose to detect the changes of mitochondrial level in hippocampal synaptosomes.The direct central mechanism of CRH and the feedback mechanism of HPA axis in the phosphorylation of tau protein and mitochondrial transport were studied by combining the antagonists of 1(CRH Receptor1CRHR1 and Glucocorticoid Receptorgrass).In order to further investigate the relationship between tau protein phosphorylation and mitochondrial transport, we used real-time imaging technique to observe the dynamic changes of mitochondrial transport when the phosphorylation level of tau protein in primary neurons changed.It was found that:1.The model of mother and child deprivation depression has typical depressive behavior and overactivated HPA axis.In open-field, sugar water preference and dark box behavioral tests, the depression model had typical depressive behavior.Compared with the control group, the depression model had significantly increased plasma cortitosterone and hypothalamic CRH mRNA levels.2.The level of phosphorylation of tau protein in hippocampus of depression model was significantly increased.Compared with the control group, there was no significant change in the total expression of tau protein in the hippocampus of the depressed rats, but it was significantly increased at the AT-8 phosphorylation site and decreased significantly at the tau-1 non-phosphorylation site.3.The level of mitochondria in hippocampal synaptosomes was significantly increased in depression model.Compared with the control group, the level of mitochondria in the hippocampal synaptosomes was significantly increased in the depression model.4.CRHR1, rather than gr, mediates increased phosphorylation of tau protein and mitochondrial level in synaptosomes.CP154526, an antagonist of CRHR1, could significantly reduce the phosphorylation of tau protein and restore mitochondrial level in synaptosomes, but RU486, the gr antagonist, did not.5.In primary neurons, when the phosphorylation level of tau protein decreased, the axoplasmic transport level of mitochondria was down-regulated.We use lithium salt to inhibit the activity of Gsk3beta and thus reduce the phosphorylation of tau protein in primary neurons.We observed a significant decrease in the axonal transport level of mitochondria.These results suggest that CRHR1 mediates the excessive phosphorylation of tau protein in the depression model and is accompanied by an increase in the mitochondrial level in the synaptosome.This reveals that the central role of CRH participates in the regulation of mitochondrial transport in depressive models, which may provide a new way to study the pathogenesis of depression.
【學位授予單位】：中國科學技術(shù)大學
【學位級別】：博士
【學位授予年份】：2011
【分類號】：R749.4;R-332
【共引文獻】
相關(guān)博士學位論文 前2條
1 楊景雷;精神分裂癥的體液代謝組學研究[D];上海交通大學;2011年
2 紀寶虎;抗精神分裂癥藥物處理大鼠的比較蛋白質(zhì)組學研究[D];上海交通大學;2009年
，
本文編號：1742455
本文鏈接：http://sikaile.net/xiyixuelunwen/1742455.html
本文選題：抑郁(Depression) + 母子剝奪(Maternal ； 參考：《中國科學技術(shù)大學》2011年博士論文
【摘要】：Tau蛋白是一種微管相關(guān)蛋白,其磷酸化受到嚴格調(diào)控,在神經(jīng)元中和軸漿運輸密切相關(guān)。據(jù)研究報道,急性應(yīng)激中,嚙齒類動物海馬中tau蛋白的磷酸化和下丘腦-垂體-腎上腺(Hypothalamo-Pituitary-Adrenal,HPA)軸的過度激活密切相關(guān)。而在抑郁病人中,HPA軸的過度激活是基本的特征之一。因此,在抑郁癥中,tau蛋白的磷酸化情況以及和軸漿運輸?shù)年P(guān)系需要進一步研究。我們的假設(shè)是:在抑郁模型中,HPA軸的過度激活會引起tau蛋白的過度磷酸化并進一步影響到軸漿運輸。 為驗證該假設(shè),我們首先建立了母子剝奪抑郁模型,對其進行了行為學測試,并檢測了HPA軸活性。然后檢測了海馬中tau蛋白的磷酸化水平。為了研究軸漿運輸?shù)淖兓?我們選擇了檢測海馬突觸體中線粒體水平的變化。結(jié)合CRH(Corticotropin-Releasing Hormone,CRH)受體1(CRH Receptor1,CRHR1)和糖皮質(zhì)激素受體(Glucocorticoid Receptor,GR)的拮抗劑,研究了CRH直接的中樞作用機制和HPA軸的反饋機制在tau蛋白磷酸化和線粒體運輸中的作用,。為了進一步檢測tau蛋白磷酸化和線粒體運輸?shù)年P(guān)系,我們用實時成像技術(shù),觀察當原代神經(jīng)元中tau蛋白磷酸化水平發(fā)生變化時線粒體運輸?shù)膭討B(tài)變化。結(jié)果發(fā)現(xiàn): 1.母子剝奪抑郁模型具有典型的抑郁樣行為和過度激活的HPA軸。 在曠場、糖水偏好和明暗箱行為學測試中,抑郁模型具有典型的抑郁樣行為。和對照組相比,抑郁模型具有顯著升高的血漿cortitosterone和下丘腦CRH mRNA水平。 2.抑郁模型海馬中tau蛋白的磷酸化水平顯著升高。 和對照組相比,抑郁鼠的海馬中tau蛋白的總表達水平?jīng)]有顯著變化,但在在AT-8磷酸化位點顯著升高,在tau-1非磷酸化位點顯著降低。 3.抑郁模型海馬突觸體中的線粒體水平顯著升高。 和對照組相比,抑郁模型海馬突觸體中的線粒體水平顯著升高。 4. CRHR1而不是GR介導了tau蛋白的磷酸化水平升高和突觸體中線粒體水平的升高。 CRHR1的拮抗劑CP154526能顯著降低tau蛋白的磷酸化,并能恢復突觸體中線粒體的水平,而GR的拮抗劑RU486卻沒有這些作用。 5.在原代神經(jīng)元中,當tau蛋白磷酸化水平降低時,線粒體的軸漿運輸水平下調(diào)。我們用鋰鹽抑制Gsk3beta的活性從而降低原代神經(jīng)元中tau蛋白的磷酸化水 平,我們觀察到線粒體的軸漿運輸水平顯著下降。 上述結(jié)果表明,在抑郁模型中CRHR1介導了tau蛋白的過度磷酸化,同時伴隨著突觸體中線粒體水平的升高。這向我們揭示,在抑郁模型中,CRH的中樞作用參與了線粒體運輸?shù)恼{(diào)控,這可能為研究抑郁發(fā)病機制提出了一條新思路。
[Abstract]:Tau protein is a microtubule-associated protein, its phosphorylation is strictly regulated, transport in neurons and axonal transport is closely related.It has been reported that the phosphorylation of tau protein in the hippocampus of rodents is closely related to the hyperactivation of the hypothalamo-Pituitary-Adrenalar axis during acute stress.Hyperactivation of HPA axis is one of the basic characteristics in depressive patients.Therefore, the phosphorylation of tau and its relationship with axonal transport in depression need further study.Our hypothesis is that excessive activation of the HPA-axis in depression models leads to excessive phosphorylation of tau proteins and further affects axonal transport.To verify this hypothesis, we first established a model of mother-son deprivation depression, tested it by behavioral tests, and measured the activity of the HPA axis.Then the phosphorylation level of tau protein in hippocampus was detected.In order to study the changes of axonal transport, we chose to detect the changes of mitochondrial level in hippocampal synaptosomes.The direct central mechanism of CRH and the feedback mechanism of HPA axis in the phosphorylation of tau protein and mitochondrial transport were studied by combining the antagonists of 1(CRH Receptor1CRHR1 and Glucocorticoid Receptorgrass).In order to further investigate the relationship between tau protein phosphorylation and mitochondrial transport, we used real-time imaging technique to observe the dynamic changes of mitochondrial transport when the phosphorylation level of tau protein in primary neurons changed.It was found that:1.The model of mother and child deprivation depression has typical depressive behavior and overactivated HPA axis.In open-field, sugar water preference and dark box behavioral tests, the depression model had typical depressive behavior.Compared with the control group, the depression model had significantly increased plasma cortitosterone and hypothalamic CRH mRNA levels.2.The level of phosphorylation of tau protein in hippocampus of depression model was significantly increased.Compared with the control group, there was no significant change in the total expression of tau protein in the hippocampus of the depressed rats, but it was significantly increased at the AT-8 phosphorylation site and decreased significantly at the tau-1 non-phosphorylation site.3.The level of mitochondria in hippocampal synaptosomes was significantly increased in depression model.Compared with the control group, the level of mitochondria in the hippocampal synaptosomes was significantly increased in the depression model.4.CRHR1, rather than gr, mediates increased phosphorylation of tau protein and mitochondrial level in synaptosomes.CP154526, an antagonist of CRHR1, could significantly reduce the phosphorylation of tau protein and restore mitochondrial level in synaptosomes, but RU486, the gr antagonist, did not.5.In primary neurons, when the phosphorylation level of tau protein decreased, the axoplasmic transport level of mitochondria was down-regulated.We use lithium salt to inhibit the activity of Gsk3beta and thus reduce the phosphorylation of tau protein in primary neurons.We observed a significant decrease in the axonal transport level of mitochondria.These results suggest that CRHR1 mediates the excessive phosphorylation of tau protein in the depression model and is accompanied by an increase in the mitochondrial level in the synaptosome.This reveals that the central role of CRH participates in the regulation of mitochondrial transport in depressive models, which may provide a new way to study the pathogenesis of depression.
【學位授予單位】：中國科學技術(shù)大學
【學位級別】：博士
【學位授予年份】：2011
【分類號】：R749.4;R-332
【共引文獻】
相關(guān)博士學位論文 前2條
1 楊景雷;精神分裂癥的體液代謝組學研究[D];上海交通大學;2011年
2 紀寶虎;抗精神分裂癥藥物處理大鼠的比較蛋白質(zhì)組學研究[D];上海交通大學;2009年
，本文編號：1742455
本文鏈接：http://sikaile.net/xiyixuelunwen/1742455.html
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