本文選題：神經(jīng)元 + 腦創(chuàng)傷��； 參考：《第四軍醫(yī)大學(xué)》2013年博士論文

【摘要】：創(chuàng)傷性腦損傷(Traumatic brain injury, TBI)是人類(lèi)致傷致殘的主要原因之一，其可分為原發(fā)性腦損傷和繼發(fā)性腦損傷，其中繼發(fā)性腦損傷是導(dǎo)致患者死亡的重要原因。TBI早期谷氨酸大量釋放，作用于各類(lèi)谷氨酸受體而發(fā)揮興奮性神經(jīng)毒作用，引起Ca2+通道持續(xù)開(kāi)放致神經(jīng)元凋亡，是加重繼發(fā)性損傷的重要原因。谷氨酸與配體門(mén)控型Ca2+通道NMDA受體結(jié)合后激活該受體，允許Ca2+大量通過(guò)，并且由于NMDAR失活緩慢,致使細(xì)胞內(nèi)Ca2+濃度升高上千倍，加重繼發(fā)性損害，因此研究如何降低NMDA受體引起的神經(jīng)元損傷，具有顯著的臨床意義。 Homer1a分子屬于Homer蛋白家族，是一個(gè)作用廣泛的即早基因(ImmediateEarly Gene)，和多種疾病的發(fā)生發(fā)展都有緊密聯(lián)系，能夠調(diào)控多種細(xì)胞分子功能。作為突觸后致密物質(zhì)（PSD）的一員，Homer1a可以調(diào)控突觸后致密物之間的信號(hào)轉(zhuǎn)導(dǎo)，是一個(gè)重要的鈣調(diào)節(jié)蛋白，它也可以通過(guò)物理替代相應(yīng)結(jié)構(gòu)起到負(fù)向調(diào)節(jié)作用，并且Homer1a能與Shank、NMDA、nNOS等分子發(fā)生相互作用。目前Homer1a與NMDAR所致?lián)p傷的關(guān)系以及和NMDAR下游分子通路的關(guān)系尚不清楚，由于NMDA受體在繼發(fā)性腦損傷中的重要作用，研究Homer1a在NMDA受體所致神經(jīng)元損傷中的作用具有深遠(yuǎn)意義。實(shí)驗(yàn)一NMDA動(dòng)物模型的建立 目的建立一種可靠而且穩(wěn)定的小鼠NMDA腦損傷模型，為進(jìn)一步研究損傷后腦組織病理生理變化及NMDA損傷機(jī)制奠定基礎(chǔ)。方法將小鼠隨機(jī)分為假手術(shù)組和NMDA皮層注射損傷組，檢測(cè)NMDA腦損傷后NSS評(píng)分和血清NSE，觀察甲苯胺藍(lán)染色變化，闡明小鼠皮層注射N(xiāo)MDA致腦損傷模型的效果。結(jié)果小鼠皮層注射N(xiāo)MDA損傷后24h，光鏡下觀察不同組甲苯胺藍(lán)染色變化發(fā)現(xiàn)：假損傷組皮層完整，尼氏體清晰可見(jiàn)，皮層無(wú)損傷，損傷組損傷灶周?chē)窠?jīng)元數(shù)量明顯減少，染色變淺，皮層完整性受到破壞。皮層注射N(xiāo)MDA后觀察小鼠行為發(fā)現(xiàn)：NMDA損傷組較假損傷組NSS評(píng)分高，說(shuō)明損傷嚴(yán)重。在損傷后12h，24h， NMDA損傷組小鼠血清中NSE高于假損傷組。結(jié)論通過(guò)檢測(cè)腦損傷后NSS評(píng)分、血清NSE和甲苯胺藍(lán)染色的變化證實(shí)，皮層注射N(xiāo)MDA致腦損傷模型具有明顯的損傷作用，是一種簡(jiǎn)單而有效的在體NMDA腦損傷模型。 實(shí)驗(yàn)二體外培養(yǎng)小鼠腦皮層神經(jīng)元NMDA損傷模型的建立 目的建立一種可靠、穩(wěn)定而且有效的離體培養(yǎng)小鼠腦皮層神經(jīng)元NMDA損傷模型。方法對(duì)培養(yǎng)成活一周的原代小鼠腦皮層神經(jīng)元進(jìn)行神經(jīng)元純度鑒定。神經(jīng)元培養(yǎng)成活后11-15d，按照Koh等的方法，進(jìn)行離體培養(yǎng)神經(jīng)元NMDA損傷。將神經(jīng)元隨機(jī)分為假損傷對(duì)照組和NMDA損傷組，通過(guò)Hoechst染色,檢測(cè)細(xì)胞內(nèi)ROS的水平以及細(xì)胞LDH釋放，明確神經(jīng)元NMDA損傷的效果。結(jié)果對(duì)培養(yǎng)的神經(jīng)元進(jìn)行NF200染色顯示：培養(yǎng)的神經(jīng)元純化率較高。神經(jīng)元在NMDA損傷后0h，各組間的LDH濃度無(wú)明顯變化；傷后6h~24h，與就假損傷對(duì)照組相比，損傷組的LDH濃度高。進(jìn)行NMDA損傷后24h，與對(duì)照組相比，細(xì)胞的凋亡顯著增加，ROS熒光亮度顯著升高。結(jié)論我們通過(guò)檢測(cè)培養(yǎng)液乳酸脫氫酶（Lactate dehydrogenase，LDH）水平、ROS染色、以及Hoechst染色的變化證明，NMDA對(duì)神經(jīng)元有確切的致傷作用。該模型是一種簡(jiǎn)單而且有效的離體皮層神經(jīng)元NMDA損傷模型，為研究NMDA所致神經(jīng)元死亡相關(guān)分子機(jī)制等提供了一個(gè)可靠、有效平臺(tái)。 實(shí)驗(yàn)三NMDA損傷（在體與離體）后Homer1a蛋白表達(dá) 目的通過(guò)在體及離體實(shí)驗(yàn)，了解NMDA腦損傷后Homer1a表達(dá)變化。方法將小鼠或細(xì)胞分為假損傷對(duì)照組和NMDA損傷組，在NMDA腦損傷模型（在體與離體）基礎(chǔ)上，采用免疫組織化學(xué)法、免疫印跡法（Western Blotting）和PCR法檢測(cè)NMDA腦損傷后Homer1a蛋白以及mRNA的變化。結(jié)果在體模型中使用NMDA注射皮層后24h，使用免疫組化染色法觀察周邊腦組織中Homer1a蛋白表達(dá)，發(fā)現(xiàn)NMDA注射組表達(dá)Homer1a蛋白的陽(yáng)性神經(jīng)元數(shù)高于對(duì)照組。在離體NMDA腦損傷模型中，用免疫印跡法檢測(cè)Homer1a蛋白在NMDA損傷后的表達(dá)變化，發(fā)現(xiàn)NMDA細(xì)胞損傷后6h至24h，Homer1a蛋白表達(dá)量明顯增加。用普通PCR方法檢測(cè)Homer1a mRNA，發(fā)現(xiàn)相比對(duì)照組，NMDA損傷組Homer1a mRNA于損傷后1h、3h表達(dá)明顯增加。結(jié)論Homer1a蛋白和mRNA水平在NMDA損傷后增加，這提示Homer1a可能在NMDA腦損傷中發(fā)揮著重要的作用，這種表達(dá)水平的增加可能與神經(jīng)元自身保護(hù)機(jī)制有關(guān)。 實(shí)驗(yàn)四NMDA損傷對(duì)Homer1a K/O小鼠的作用 目的通過(guò)在體實(shí)驗(yàn)，明確NMDA損傷對(duì)Homer1a K/O小鼠的作用。方法首先對(duì)Homer1a基因敲除鼠進(jìn)行篩選與鑒定，后將小鼠分為2個(gè)組，KO（Knock Out）小鼠組和WT（Wild Type）小鼠組，建立小鼠NMDA腦損傷模型，用甲苯胺藍(lán)染色計(jì)算梗死面積、對(duì)小鼠進(jìn)行神經(jīng)功能學(xué)評(píng)分（NSS），以及檢測(cè)血清NSE，明確Homer1aK/O小鼠和WT小鼠的對(duì)NMDA損傷作用效應(yīng)的區(qū)別。結(jié)果我們成功雜交出了Homer1a基因敲除鼠，普通PCR結(jié)果顯示，基因敲除小鼠有效，無(wú)Homer1a基因表達(dá)，并且有Homer1c基因表達(dá)。小鼠皮層注射N(xiāo)MDA后24h，尼氏染色計(jì)算梗死面積后發(fā)現(xiàn)Homer1a K/O組較WT組腦皮層損傷重；NSS評(píng)分提示在12h、24h，Homer1aK/O組較WT組評(píng)分高；血清NSE含量檢測(cè)提示損傷后24h，Homer1a K/O組血清中NSE較WT組高。結(jié)論這些結(jié)果提示，Homer1a基因敲除加重了NMDA引起的腦損傷，Homer1a基因?qū)MDA腦損傷具有保護(hù)作用。 實(shí)驗(yàn)五體外模型中過(guò)表達(dá)Homer1a對(duì)NMDA神經(jīng)元損傷的影響 目的前面的實(shí)驗(yàn)結(jié)果提示，敲除小鼠Homer1a基因加重NMDA腦損傷，推論Homer1a基因?qū)MDA腦損傷有保護(hù)作用，但仍需進(jìn)一步細(xì)胞模型中得到證實(shí)。方法進(jìn)行腦皮層神經(jīng)元原代培養(yǎng)后，轉(zhuǎn)染LV-Homer1a，鑒定表達(dá)后，將細(xì)胞分為對(duì)照損傷組，空載體組，Homer1a過(guò)表達(dá)組，建立NMDA損傷模型，觀察細(xì)胞細(xì)胞死亡率，進(jìn)行Hoechst染色分析以及損傷后LDH值測(cè)定，Western-Blot進(jìn)行損傷后p-Caspase-3分析。結(jié)果通過(guò)使用過(guò)表達(dá)Homer1a的慢病毒載體（LV-Homer1a）轉(zhuǎn)染神經(jīng)元發(fā)現(xiàn)過(guò)表達(dá)Homer1a成功，能夠明顯檢測(cè)出外源性的Homer1a。用慢病毒載體（LV-Homer1a）轉(zhuǎn)染神經(jīng)元，造模后24h，對(duì)各組神經(jīng)元進(jìn)行Hoechst染色，發(fā)現(xiàn)轉(zhuǎn)染LV-Homer1a降低了凋亡細(xì)胞比例和，降低了p-Caspase3表達(dá)。LDH結(jié)果提示傷后6h、12h、24h，與對(duì)照損傷組、LV-Vector損傷組比，LV-Homer1a的LDH濃度較低。結(jié)論這些結(jié)果提示，離體試驗(yàn)中，Homer1a能夠降低凋亡細(xì)胞的比例，保護(hù)神經(jīng)元，也驗(yàn)證了在體試驗(yàn)的結(jié)果。 實(shí)驗(yàn)六Homer1a對(duì)NMDAR致?lián)p傷功能的影響以及nNOS的活性影響 目的前面的實(shí)驗(yàn)結(jié)果證實(shí)，Homer1a對(duì)NMDA所致?lián)p傷后的神經(jīng)元具有直接保護(hù)作用，但具體的機(jī)制尚不清楚。本實(shí)驗(yàn)擬在離體模型中研究損傷后Homer1a對(duì)NMDAR下游通路的影響，以及Homer1a對(duì)nNOS活性的影響。方法腦皮層神經(jīng)元原代培養(yǎng)成活后轉(zhuǎn)染LV-Homer1a，建立NMDA損傷模型，隨機(jī)將細(xì)胞分為NMDA損傷組、空載體組、轉(zhuǎn)染LV-Homer1a組，觀察細(xì)胞內(nèi)產(chǎn)生的ROS，Ca2+、并進(jìn)行全細(xì)胞膜片鉗記錄，用Western Blot檢測(cè)各組中細(xì)胞內(nèi)p-nNOS、 p-ERK、 p-CREB的表達(dá)。結(jié)果神經(jīng)元NMDA損傷后24h，與對(duì)照組相比，轉(zhuǎn)染LV-Homer1a降低了ROS生成和NMDA引起的鈣離子內(nèi)流。轉(zhuǎn)染LV-Homer1a降低了神經(jīng)元NMDA受體的電流峰值。Western Blot結(jié)果提示：轉(zhuǎn)染LV-Homer1a后降低NMDAR過(guò)度激活導(dǎo)致的ERK，CREB，nNOS的激活。結(jié)論這些結(jié)果提示，離體神經(jīng)元轉(zhuǎn)染LV-Homer1a后，改變了NMDAR的屬性，降低了其通透性，減弱其下游通路的過(guò)度激活，這為進(jìn)一步闡明了Homer1a蛋白對(duì)NMDA腦損傷的保護(hù)機(jī)制奠定了基礎(chǔ)。 實(shí)驗(yàn)七Homer1a對(duì)神經(jīng)元中NMDAR以及NMDAR復(fù)合體的影響 目的前面的實(shí)驗(yàn)結(jié)果證實(shí)，Homer1a對(duì)NMDA所致神經(jīng)元損傷具有直接保護(hù)作用，且改變了NMDA屬性以及其下游通路的活性，但是具體的機(jī)制尚不清楚。我們推測(cè)，Homer1a可能是通過(guò)調(diào)控NMDAR本身分布以及NMDAR和相關(guān)的分子集團(tuán)結(jié)合發(fā)揮保護(hù)作用。方法通過(guò)向HEK293T細(xì)胞轉(zhuǎn)染NR1NR2B受體以及Homer1a，進(jìn)行膜片鉗記錄，按照轉(zhuǎn)染的分子不同分為空載體組、NR1/NR2B組、NR1/NR2B/Con組、NR1/NR2B/Homer1a組，研究Homer1a對(duì)NMDA受體功能的直接影響。在神經(jīng)元轉(zhuǎn)染LV-Homer1a后，將細(xì)胞分為空載體組、過(guò)表達(dá)Homer1a組，通過(guò)提取提純膜蛋白，，檢測(cè)Homer1a對(duì)NMDA受體亞基NR2B分布的影響，通過(guò)免疫共沉淀，檢測(cè)轉(zhuǎn)染LV-Homer1a對(duì)NMDA受體亞基NR2B與nNOS蛋白結(jié)合的影響。結(jié)果在293T細(xì)胞上我們成功過(guò)表達(dá)了NR1NR2B，且具有受體活性，同時(shí)轉(zhuǎn)染LV-Homer1a發(fā)現(xiàn)，Homer1a對(duì)293T細(xì)胞上的NR1/NR2B受體的峰值無(wú)明顯影響。體外培養(yǎng)神經(jīng)元轉(zhuǎn)染LV-Homer1a72h，發(fā)現(xiàn)轉(zhuǎn)染LV-Homer1a后降低膜上NR2B分布。通過(guò)對(duì)NR2B以及nNOS進(jìn)行免疫沉淀發(fā)現(xiàn)，轉(zhuǎn)染LV-Homer1a降低了NR2B和nNOS之間結(jié)合。 結(jié)論這些結(jié)果提示，Homer1a能夠促進(jìn)NMDAR亞基NR2B的內(nèi)陷以及破壞NR2B,nNOS之間的結(jié)合，這兩點(diǎn)可能是Homer1a降低NMDA神經(jīng)元損傷的重要機(jī)理。
[Abstract]:Traumatic brain injury (TBI) is one of the main causes of human disability, which can be divided into primary brain injury and secondary brain injury. Secondary brain injury is an important cause of death in patients with.TBI release of glutamic acid at early stage, acting on all kinds of glutamate receptors and exerting excitatory neurotoxicity. The sustained and open neuronal apoptosis of the Ca2+ channel is an important cause of secondary injury. The binding of glutamic acid to the ligand gated Ca2+ channel NMDA receptor activates the receptor, allowing the Ca2+ to pass through, and the increase of the concentration of Ca2+ in the cells increases thousands of times and aggravates secondary damage due to the slow inactivation of NMDAR, thus how to decrease the study. NMDA receptor induced neuronal damage has significant clinical significance.
Homer1a is a family of Homer protein, which is a widely used ImmediateEarly Gene, which is closely related to the occurrence and development of various diseases and can regulate the function of many kinds of cell molecules. As a member of the postsynaptic dense substance (PSD), Homer1a can regulate the signal transduction between the postsynaptic densification and is an important factor. The calcium regulatory protein can also play a negative regulatory role by replacing the corresponding structure by physics, and Homer1a can interact with Shank, NMDA, nNOS and other molecules. The relationship between Homer1a and NMDAR induced damage and the relationship with the downstream pathway of the downstream NMDAR is unclear, due to the important role of NMDA receptor in secondary brain damage. It is of great significance to study the role of Homer1a in NMDA receptor induced neuronal damage. Experiment 1 NMDA animal model was established.
Objective to establish a reliable and stable model of NMDA brain injury in mice, and to lay the foundation for further study of the pathophysiological changes of brain tissue and the mechanism of NMDA injury after injury. Methods mice were randomly divided into sham operation group and NMDA cortex injuried group. The NSS score and serum NSE were detected after NMDA brain injury, and the changes of toluidine blue were observed. The effect of NMDA induced brain injury in the cortex of the mice was given. Results the cortex was injected with 24h after NMDA injury. The changes of toluidine blue in different groups were observed under light microscope: the cortex of the false injury group was complete, the Nissl body was clearly visible, the cortex was not damaged, the number of neurons around the lesion was significantly reduced, the staining was shallow, and the cortex integrity was affected. After the injury, the mouse behavior was observed after the cortex injection of NMDA: the NMDA injury group was higher than the false injury group, which indicated that the injury was serious. The serum NSE of the mice in the 12h, 24h, NMDA injury group was higher than that of the false injury group after the injury. Conclusion the changes of NSS score after the detection of brain injury, the changes of serum NSE and toluidine blue staining proved that the cortex injection of NMDA caused brain damage. It is a simple and effective in vivo NMDA brain injury model.
Experiment two establishment of NMDA damage model of mouse cortical neurons in vitro
Objective to establish a reliable, stable and effective NMDA damage model for the cultured mouse cortical neurons in vitro. Methods the neuron purity of the primary mouse cortical neurons cultured for one week was identified. The neuron culture was 11-15d, and the neuron NMDA damage in vitro was carried out according to the method of Koh. The neurons were randomly divided. For the false injury control group and the NMDA injury group, the level of ROS in the cells and the release of LDH were detected by Hoechst staining. The effect of NMDA damage on the neurons was determined. Results the cultured neurons were stained with NF200 to show that the cultured neurons had higher purification rate. There was no obvious change in LDH concentration in each group after the injury of NMDA. After 6h~24h, the concentration of LDH in the injured group was higher than that of the control group. After NMDA injury, 24h, compared with the control group, the apoptosis of the cells increased significantly and the ROS fluorescence luminance increased significantly. Conclusion we have proved that the NMDA pairs were detected by detecting the level of Lactate dehydrogenase (LDH), ROS staining, and Hoechst staining. This model is a simple and effective model of NMDA damage in isolated cortical neurons, which provides a reliable and effective platform for the study of the molecular mechanism of neuronal death induced by NMDA.
Experiment three the expression of Homer1a protein after NMDA injury (in vitro and in vivo).
Objective to understand the changes of Homer1a expression after NMDA brain injury in vivo and in vitro. Methods mice or cells were divided into false damage control group and NMDA injury group. On the basis of NMDA brain damage model (in vivo and in vitro), immunohistochemistry, Western Blotting and PCR method were used to detect the Homer1a protein after NMDA brain injury. Results in the body model, 24h was injected after the injection of NMDA in the body model, and the expression of Homer1a protein in the peripheral brain tissue was observed by immunohistochemical staining. The number of positive neurons expressing Homer1a protein in NMDA injection group was higher than that of the control group. In the isolated NMDA brain injury model, the immunoblotting method was used to detect the Homer1a protein after NMDA injury. The expression of 6h to 24h and the expression of Homer1a protein increased significantly after NMDA cell injury. Homer1a mRNA was detected by common PCR method. It was found that the Homer1a mRNA of NMDA damage group was 1h after injury, and the 3H expression increased obviously. This increase in expression level may be related to the mechanism of neuronal self protection.
Effect of Experiment four NMDA damage on Homer1a K/O mice
Objective to determine the effect of NMDA injury on Homer1a K/O mice in vivo. Methods first of all, the Homer1a gene knockout mice were screened and identified. Then the mice were divided into 2 groups, KO (Knock Out) mice and WT (Wild Type) mice. The mouse NMDA brain damage model was established. The infarct area was calculated with toluidine blue staining, and the mice were nervous. The functional score (NSS), and the detection of serum NSE, make clear the difference between the Homer1aK/O mice and the WT mice. Results we successfully hybridized the Homer1a gene knockout mice. The common PCR results showed that the gene knockout mice were effective, no Homer1a gene expression, and the Homer1c gene expression. The mouse cortex was injected with NMDA 24h, Nei. It was found that Homer1a K/O group was more severe than group WT, and NSS score was higher in 12h, 24h, Homer1aK/O group than that in group WT, and the serum NSE content detection suggested that the serum levels of 24h and Homer1a K/O group were higher than those in the group of WT. Conclusion these results suggest that the gene knockout aggravates the brain damage caused by the gene knockout. The gene has protective effects on NMDA brain damage.
Effect of overexpression of Homer1a in vitro on NMDA neuron injury in experiment five
The previous experimental results suggest that the Homer1a gene in the knockout mice aggravates NMDA brain damage and that the Homer1a gene has a protective effect on the brain damage of NMDA, but it still needs to be confirmed in the further cell model. After the primary culture of the cortical neurons, the transfected LV-Homer1a was divided into the control injury group and the empty body group after the transfection of LV-Homer1a. Homer1a overexpression group, establish NMDA damage model, observe cell death rate, analyze Hoechst staining and determine LDH value after injury, p-Caspase-3 analysis after Western-Blot damage. The result can be detected by transfection of Homer1a based lentivirus vector (LV-Homer1a) to the expression of Homer1a, which can be detected obviously. Exogenous Homer1a. transfected neurons with lentivirus vector (LV-Homer1a), and then 24h, Hoechst staining was performed on each group of neurons. It was found that transfection of LV-Homer1a reduced the proportion of apoptotic cells and decreased the p-Caspase3 expression.LDH results, suggesting 6h, 12h, 24h, compared with the control injury group, LV-Vector injury group, and low concentration of LV-Homer1a. Conclusion these results suggest that Homer1a can reduce the proportion of apoptotic cells and protect neurons in vitro, and also verify the results of in vivo experiments.
Experiment six the effect of Homer1a on NMDAR induced damage and the activity of nNOS.
The previous experimental results confirm that Homer1a has a direct protective effect on the neurons injured by NMDA, but the specific mechanism is still unclear. This experiment is intended to study the effect of Homer1a on the downstream pathway of NMDAR after injury and the effect of Homer1a on the activity of nNOS. LV-Homer1a, the NMDA damage model was established, and the cells were randomly divided into NMDA injury group, empty body group, transfected LV-Homer1a group, and observed the ROS, Ca2+, and whole cell patch clamp recording. The expression of p-nNOS, p-ERK, p-CREB in each group was detected by Western Blot. Results the neuron NMDA was injured and transfected compared with the control group. LV-Homer1a reduced the influx of calcium ions caused by ROS formation and NMDA. Transfection of LV-Homer1a reduced the peak current peak of NMDA receptor.Western Blot results, suggesting that the transfection of LV-Homer1a to reduce ERK, CREB, nNOS activation caused by NMDAR overactivation. The property, which reduces its permeability and weakens the over activation of its downstream pathway, lays the foundation for further clarifying the protective mechanism of Homer1a protein to NMDA brain damage.
Effect of experiment seven Homer1a on NMDAR and NMDAR complex in neurons
The previous experimental results confirm that Homer1a has a direct protective effect on NMDA induced neuronal damage and changes the NMDA property and its downstream pathway activity, but the specific mechanism is still unclear. We speculate that Homer1a may play a protective role by regulating the distribution of NMDAR itself and the combination of NMDAR and related molecular groups. Methods through the transfection of NR1NR2B receptor and Homer1a to HEK293T cells, the patch clamp recording was carried out. According to the different transfection molecules, the cells were divided into empty carrier group, NR1/NR2B group, NR1/NR2B/Con group and NR1/NR2B/Homer1a group, to study the direct effect of Homer1a on the function of NMDA receptor. After transfection of LV-Homer1a, the cells were divided into empty carrier group and overexpressed Homer. In group 1a, the effect of Homer1a on the distribution of NMDA receptor subunit NR2B was detected by extracting purified membrane protein, and the effect of LV-Homer1a on the binding of NMDA receptor subunit NR2B to nNOS protein was detected by immunoprecipitation. The results showed that NR1NR2B was successfully expressed on 293T cells, with receptor activity, and LV-Homer1a discovery was transfected, Homer1a against 29 The peak value of NR1/NR2B receptor on 3T cells was not significantly affected. The transfection of LV-Homer1a72h to neurons in vitro was found to reduce the distribution of NR2B on the membrane. By immunoprecipitation of NR2B and nNOS, the transfection of LV-Homer1a reduced the binding between NR2B and nNOS.
Conclusion these results suggest that Homer1a can promote the collapse of NMDAR subunit NR2B and destroy the binding of NR2B and nNOS, which may be an important mechanism for Homer1a to reduce the damage of NMDA neurons.
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R651.15

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