本文選題：CREB　切入點(diǎn)：杏仁體　出處：《復(fù)旦大學(xué)》2005年博士論文　論文類型：學(xué)位論文

【摘要】：杏仁體(amygdala, AM)和海馬結(jié)構(gòu)(hippocampal formation, HF)是與情緒活動(dòng)和學(xué)習(xí)記憶相關(guān)的重要結(jié)構(gòu),AM和HF內(nèi)的轉(zhuǎn)錄因子cAMP反應(yīng)元件結(jié)合蛋白(CREB)被證實(shí)在記憶調(diào)節(jié)過(guò)程中發(fā)揮了重要作用。CREB是一種核蛋白,是刺激誘導(dǎo)轉(zhuǎn)錄因子的主要成員,多種刺激都可導(dǎo)致CREB的活化,形成磷酸化的CREB(pCREB)。神經(jīng)生理及神經(jīng)藥理學(xué)的研究表明,在突觸可塑性的長(zhǎng)時(shí)程效應(yīng)階段,CREB可調(diào)節(jié)基因轉(zhuǎn)錄、參與蛋白質(zhì)合成,從而維持突觸效率的長(zhǎng)期持續(xù)改變。但是形態(tài)上,目前還不清楚AM、HF等腦區(qū)內(nèi)的CREB究竟在哪類神經(jīng)元中表達(dá)而發(fā)揮作用。為此本實(shí)驗(yàn)準(zhǔn)備進(jìn)一步探討:① 應(yīng)激刺激后AM和HF內(nèi)pCREB表達(dá)量的變化;② pCREB在什么神經(jīng)元中表達(dá);③ pCREB的表達(dá)與NMDA受體活性的關(guān)系。本實(shí)驗(yàn)作了如下工作: 1.對(duì)SD大鼠強(qiáng)迫性游泳(FS),建立情緒性應(yīng)激刺激動(dòng)物模型,以正常同類大鼠為對(duì)照,用免疫細(xì)胞化學(xué)單標(biāo)法、蛋白免疫印跡實(shí)驗(yàn),以抗pCREB抗體觀察檢測(cè)pCREB免疫陽(yáng)性神經(jīng)元在對(duì)照組和實(shí)驗(yàn)組大鼠AM各亞核團(tuán)和HF內(nèi)的分布特征及變化規(guī)律;用免疫細(xì)胞化學(xué)雙標(biāo)法和免疫電鏡,以抗pCREB、抗Glu和抗PV抗體,分析了表達(dá)pCREB神經(jīng)元的類型。結(jié)果顯示:①FS刺激后,pCREB免疫陽(yáng)性細(xì)胞核在AM的外測(cè)核(LA)、基底外側(cè)核(BL)、中央核(Ce)以及HF的齒狀回(DG)和CA3區(qū)的數(shù)量明顯增加。FS 1h后Ce內(nèi)pCREB陽(yáng)性細(xì)胞核數(shù)目最多,為1082±229個(gè)/mm~2;而LA內(nèi)的pCREB陽(yáng)性細(xì)胞核數(shù)目變化最大,對(duì)照組為578±106個(gè)/mm~2;而FS 15min后則為1078±338個(gè)/mm~2。在DG顆粒細(xì)胞層內(nèi),對(duì)照組的pCREB陽(yáng)性細(xì)胞核數(shù)目很少,成線狀散在分布;而FS后pCREB陽(yáng)性細(xì)胞核的數(shù)目明顯增多,成帶狀密布于整個(gè)DG顆粒細(xì)胞層。FS后,AM各亞核團(tuán)內(nèi)pCREB免疫陽(yáng)性細(xì)胞核的變化趨勢(shì)具有時(shí)間相關(guān)性,本實(shí)驗(yàn)觀察到,LA內(nèi)的pCREB表達(dá)量在FS后15min達(dá)到高峰,而Ce內(nèi)的pCREB則在FS后1h表達(dá)量最高。免疫蛋白印跡的實(shí)驗(yàn)結(jié)果與上述變化密切相關(guān),顯示,FS刺激后AM各亞核和HF內(nèi)pCREB總蛋白量都明顯增加。②免疫細(xì)胞化學(xué)雙標(biāo)結(jié)果顯示,在LA、BL、Ce、DG和CA3區(qū)中,呈綠色熒光的pCREB免疫陽(yáng)性神經(jīng)元細(xì)胞核,位于呈紅色Glu免疫陽(yáng)性的神經(jīng)元胞質(zhì)內(nèi),而未見(jiàn)其在
[Abstract]:Amygdala (AM) and hippocampal formation (HFF) are important structures associated with emotional activity and learning and memory. The transcription factor cAMP response element binding protein (cAMP) in HF has been proved to play an important role in memory regulation. CREB is a nuclear protein. Multiple stimuli can activate CREB and form phosphorylated CREBPCREB.Neurophysiological and neuropharmacological studies have shown that CREB can regulate gene transcription during the long term effect of synaptic plasticity. Participate in protein synthesis, thereby sustaining long-term changes in synaptic efficiency. But morphologically, It is not clear what kind of neurons CREB is expressed in the brain regions such as AMN / HF. Therefore, this study is going to further explore the changes of pCREB expression in AM and HF after the stress of 1: 1 in which neurons do the 2 pCREB express? The relationship between the expression of pCREB and the activity of NMDA receptor. 1. The animal model of emotional stress stimulation was established in SD rats by compulsive swimming. The normal rats were used as control, immunocytochemical single labeling method and Western blot were used. The distribution and changes of pCREB immunoreactive neurons in the subnuclei and HF of AM in the control group and experimental group were observed and observed by using anti pCREB antibody, and the immunocytochemical double labeling method and immunoelectron microscope were used to detect the distribution of pCREB immunoreactive neurons in the control group and the experimental group, and the anti-pCREBand anti Glu and anti PV antibodies were used. The expression of pCREB neurons was analyzed. The results showed that the number of pCREB immunoreactive nuclei in the outer nucleus of AM, the basolateral nucleus, the central nucleus, and the number of the dentate gyrus DGG) and the CA3 region of HF increased significantly. FS 1 h after stimulation, the number of pCREB in ce was significantly increased. The number of positive nuclei is the highest. The number of pCREB positive nuclei in LA was the highest, 578 鹵106 / mm2 in control group and 1078 鹵338 / mm-2 in DG granular cell layer 15 minutes after FS. In DG granular cell layer, the number of pCREB positive nuclei in control group was very small and distributed linearly. However, the number of pCREB positive nuclei increased significantly after FS, and the change trend of pCREB immunoreactive nuclei in the whole DG granular cell layer. FS was time-dependent. In this study, we observed that the expression of pCREB in LA reached its peak at 15 minutes after FS, while the expression of pCREB in ce was the highest at 1 hour after FS. The results of Western blot were closely related to the above changes. The results showed that the total protein content of pCREB in AM subnuclei and HF increased significantly after FS-stimulation. The results showed that in the DG and CA3 regions of BLCE-DG and CA3, the nuclei of pCREB immunoreactive neurons were green fluorescent. Located in the cytoplasm of red Glu immunoreactive neurons, but not found in the
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R329

【引證文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 林棟;針刺大鼠外關(guān)穴對(duì)不同腦區(qū)CREB磷酸化及相關(guān)信號(hào)轉(zhuǎn)導(dǎo)機(jī)制研究[D];廣州中醫(yī)藥大學(xué);2011年

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本文編號(hào)：1613935