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δ-catenin,spinophilin在大鼠小腦的表達(dá)及其年齡差異

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  本文關(guān)鍵詞: δ-連環(huán)素 樹突棘素 小腦 年齡 大鼠 出處:《華中科技大學(xué)》2006年碩士論文 論文類型:學(xué)位論文


【摘要】: 神經(jīng)老化是生命活動(dòng)的必然,探討神經(jīng)老化機(jī)制,尋找延緩神經(jīng)老化進(jìn)程的方法是神經(jīng)科學(xué)領(lǐng)域里的一個(gè)重要內(nèi)容。在腦衰老研究中,突觸可塑性備受人們關(guān)注。δ-連環(huán)素(δ-catenin)與樹突棘素(Spinophilin)是近年來發(fā)現(xiàn)的分布于神經(jīng)元樹突的蛋白,在突觸塑形中扮演重要角色。δ-catenin是p120-catenin (p120ctn)蛋白家族成員,具有能和鈣粘素(cadherins)近膜端結(jié)合的序列,δ-catenin是這一蛋白家族唯一特異性只表達(dá)于神經(jīng)系統(tǒng)的蛋白,它能和突觸后成分蛋白的PDZ結(jié)構(gòu)域結(jié)合。δ-catenin定位于突觸后區(qū)域,能調(diào)節(jié)突觸的傳入和塑形。Spinophilin因其特異性定位于樹突棘頭部,故得名樹突棘素。它具有細(xì)胞連接處絞手架蛋白(scaffold protein)的特征,并且含有和PSD95/DLG/zo-1相同的序列,是蛋白磷酸酶-1(PP1)新的靶向亞單位,能將PP1定位于樹突棘的底物(如受PP1調(diào)節(jié),從而影響突觸功能的神經(jīng)遞質(zhì)受體)上,調(diào)節(jié)樹突棘的形態(tài),完成突觸塑形。小腦是腦衰老機(jī)理研究的理想模型系統(tǒng)之一。 本實(shí)驗(yàn)在建立大鼠自然老化模型的基礎(chǔ)上,運(yùn)用免疫組織化學(xué)方法檢測(cè)δ-catenin及spinophilin在不同年齡大鼠小腦的定位表達(dá);運(yùn)用圖像分析系統(tǒng)和免疫印跡(Western-blot)技術(shù)檢測(cè)δ-catenin及spinophilin在大鼠小腦的表達(dá)水平;運(yùn)用RT-PCR技術(shù)檢測(cè)δ-catenin,spinophilin mRNA在不同年齡組大鼠小腦表達(dá)的差異。 免疫組織化學(xué)檢測(cè)顯示δ-catenin,spinophilin主要表達(dá)在小腦分子層和顆粒層,在中、青年大鼠蒲肯野細(xì)胞有中等陽性表達(dá),老年大鼠蒲肯野細(xì)胞表達(dá)較弱。Western blot檢測(cè)顯示δ-catenin、spinophilin在中年組大鼠小腦表達(dá)最高,在老年組大鼠小腦表達(dá)最低。RT-PCR檢測(cè)顯示δ-catenin、spinophilin mRNA表達(dá)水平的增齡性變化與其蛋白表達(dá)的增齡性變化一致。 本實(shí)驗(yàn)研究結(jié)果表明δ-catenin,spinophilin在大鼠小腦的表達(dá)存在增齡性變化。這一變化和不同年齡大鼠小腦突觸的數(shù)量,結(jié)構(gòu)和功能的變化有關(guān)。
[Abstract]:Nerve aging is a necessity of life activity. It is an important content in the field of neuroscience to explore the mechanism of nerve aging and to find ways to delay the process of nerve aging. Synaptic plasticity has attracted much attention. 未 -catenin (未 -catenin) and spinophilin (Spinophilin) are proteins found in recent years. 未 -catenin is a member of p120-catenin / p120ctnprotein family. 未 -catenin is the only protein expressed specifically in the nervous system with a sequence that binds to the proximal end of the cadherins. It can bind to the PDZ domain of the postsynaptic component protein. 未 -catenin is located in the postsynaptic region. It can regulate the afferent and shape of synapses. Spinophilin is located in the head of dendritic spine because of its specificity. It has the characteristics of scaffold protein (scaffold protein) and contains the same sequence as PSD95/DLG/zo-1. It is a new target subunit of protein phosphatase (PP-1), which can locate PP1 on the substrate of dendritic spine (such as neurotransmitter receptor, which is regulated by PP1, which affects synaptic function). The cerebellum is one of the ideal model systems for the study of brain aging mechanism. The expression of 未 -catenin and spinophilin in cerebellum of rats of different ages was detected by immunohistochemical method on the basis of establishing natural aging model of rats. The expression of 未 -catenin and spinophilin in cerebellum of rats was detected by image analysis system and Western-blot technique. The expression of 未 -cateninine spinophilin mRNA in the cerebellum of rats of different age groups was detected by RT-PCR technique. Immunohistochemical examination showed that 未 -cateninine spinophilin was mainly expressed in the molecular and granular layers of cerebellum, and the expression of 未 -cateninine spinophilin was moderately positive in the young rats' Purkinje cells. The expression of 未 -catenin spinophilin was the highest in the middle aged rats. The lowest expression of 未 -catenin was detected by RT-PCR in the cerebellum of the aged rats. The age change of spinophilin mRNA expression level was consistent with that of its protein expression. The results showed that the expression of 未 -cateninine spinophilin in the cerebellum of rats had age-related changes and the number of cerebellar synapses in rats of different ages. Changes in structure and function are relevant.
【學(xué)位授予單位】:華中科技大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2006
【分類號(hào)】:R329

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