本文關(guān)鍵詞： 小腦皮質(zhì) 組織發(fā)生 發(fā)育 神經(jīng)元凋亡 關(guān)系　出處：《河南大學》2007年碩士論文　論文類型：學位論文

【摘要】： 小腦位于腦橋和延髓的背面及大腦枕葉的下方,是中樞神經(jīng)系統(tǒng)中與執(zhí)行運動機能有關(guān)的較高級調(diào)節(jié)中樞,并且還參與了學習記憶的神經(jīng)活動過程。成熟小腦皮質(zhì)有三層結(jié)構(gòu),包括五種神經(jīng)元,在這些神經(jīng)元中最具有特征性的是巨大的Purkinje細胞和較小的顆粒細胞,Purkinje細胞是小腦皮質(zhì)的主要神經(jīng)元,顆粒細胞是小腦數(shù)量最多的神經(jīng)元。 目前人們對小腦形態(tài)、細胞化學構(gòu)筑、纖維聯(lián)系、神經(jīng)遞質(zhì)及不同神經(jīng)元發(fā)生凋亡的可能機制等方面做了較為深入的研究。但有關(guān)小腦皮質(zhì)的具體形態(tài)演變過程及發(fā)育中的神經(jīng)元凋亡的變化規(guī)律及可能發(fā)生的機制的資料相對較少,所以此問題的闡清將更進一步加深對神經(jīng)系統(tǒng)發(fā)育過程規(guī)律的認識,也為胚胎小腦移植和某些退化性神經(jīng)系統(tǒng)疾病提供必要的幫助。 為了探討小鼠小腦皮質(zhì)的組織發(fā)生過程和發(fā)育中神經(jīng)元凋亡的規(guī)律和機制及兩者之間的關(guān)系,我們應(yīng)用光鏡和電鏡技術(shù)對胚胎和生后小腦皮質(zhì)進行形態(tài)學觀察,對生后第2天(P2)至成年鼠小腦皮質(zhì)各層厚度和細胞密度行形態(tài)學測量;用激活型Caspase-3多抗免疫組化標記及Hoechst33258染色液染色檢測從出生至成年小鼠小腦皮質(zhì)中神經(jīng)元的凋亡,用Western blotting方法對小腦組織中Caspase-3和Caspase-8的活化片段進行半定量測定。結(jié)果顯示:①胚胎12d(E12)小腦原基有室管膜層、套層和邊緣層構(gòu)成;②約出生當日(P0)出現(xiàn)較典型外顆粒層、分子層、Purkinje細胞層和內(nèi)顆粒層;③外顆粒層P6/7達最厚,至P20消失;④P0至P30,分子層原基呈CUB型曲線增厚,細胞密度呈S型曲線降低,內(nèi)顆粒層厚度呈S型曲線增加,細胞逐步分化發(fā)育成熟,密度呈CUB型曲線先增加后降低,Purkinje細胞體積呈INV型曲線增大,單位長度細胞密度呈S型曲線降低,樹突樹逐漸形成,約P7時Purkinje細胞排列成單層;其發(fā)育過程中的差異有統(tǒng)計學意義,P0.001;⑤免疫組化:外顆粒層、Purkinje細胞層和內(nèi)顆粒層凋亡細胞密度最高分別在出生后第8天(P8)、P5及P9左右,約P20各層細胞凋亡密度都很低;⑥Western blotting: Caspase-3活化片段的表達量在P5最高,以后漸降低至P14消失;Caspase-8活化片段的表達量從P0到P10都較高,以后略降低至P30基本消失;其發(fā)育過程中神經(jīng)元凋亡的差異有統(tǒng)計學意義,P0.01。 研究結(jié)果表明:E12～P0,片層化的小腦主要經(jīng)歷了細胞增殖、分化與遷移;P0～P30,片層化的小腦逐漸發(fā)育成熟,其中外顆粒層的消亡以細胞遷移和凋亡為主,而其他各層結(jié)構(gòu)主要經(jīng)歷了細胞的分化、發(fā)育與凋亡。P0～P14是小腦皮質(zhì)神經(jīng)元迅速發(fā)育時期,同時也是其凋亡發(fā)生的高峰時期,通過啟動Caspase-8的活化進而激活效應(yīng)性Caspase-3的細胞凋亡途徑存在于小腦皮質(zhì)塑型成熟過程中。
[Abstract]:The cerebellum is located at the back of the pons and medulla oblongata and below the occipital lobe of the brain. It is a higher regulatory center in the central nervous system related to the execution of motor functions, and is also involved in the neural processes of learning and memory. The mature cerebellar cortex has a three-layer structure. Among the five kinds of neurons, the large Purkinje cells and the smaller granular cells are the main neurons in the cerebellar cortex, and the granulosa cells are the most numerous neurons in the cerebellum. At present, people are concerned about the cerebellar morphology, cytochemical architecture, fiber connection, Neurotransmitters and the possible mechanisms of apoptosis of different neurons have been studied in depth. However, the specific morphologic evolution of cerebellar cortex, the changes of neuronal apoptosis in development and the possible mechanism of apoptosis have been studied. There is relatively little data available. Therefore, the explanation of this problem will further deepen the understanding of the rules of nervous system development, and also provide necessary help for embryo cerebellum transplantation and some degenerative nervous system diseases. In order to investigate the histogenesis process of mouse cerebellar cortex, the regularity and mechanism of neuronal apoptosis during development and the relationship between them, we observed the morphology of embryonic and postnatal cerebellar cortex by light microscope and electron microscope. The thickness and cell density of cerebellar cortex from the second day after birth to adult rats were measured by morphology, and the apoptosis of neurons in cerebellar cortex of mice from birth to adult was detected by immunohistochemical staining of activated Caspase-3 and Hoechst33258 staining. The activation fragments of Caspase-3 and Caspase-8 in cerebellar tissue were determined by Western blotting method. The results showed that the primordium of cerebellum had ependymal layer, and the lamina and margin of cerebellum formed a typical outer granular layer (P0). The P6 / 7 of P6 / 7 was the thickest in the cell layer of Purkinje and the outer layer of P6 / 7 in the inner granular layer. By the time P20 disappeared from 4P0 to P30, the primordium of the molecular layer was thickened by CUB curve, the cell density was decreased by S-type curve, and the thickness of the inner granular layer was increased by S-type curve. The cell density increased firstly and then decreased. The cell volume increased as INV curve and the cell density per unit length decreased as S-shaped curve. Dendritic tree was gradually formed and Purkinje cells were arranged as monolayers at P7. The difference in development process was statistically significant (P 0.001): the density of apoptotic cells in Purkinje cell layer and inner granular layer was the highest at the 8th day after birth, and the density of apoptotic cells in the outer granular layer was about P8 + P5 and P9, respectively. The apoptotic density of P20 cells was very low. The expression of Caspase-3 activation fragment was the highest in P5, then decreased to P14 disappeared. The expression of Caspase-8 activated fragment was higher from P0 to P10, then decreased slightly to P30. The difference of neuronal apoptosis during its development was statistically significant (P 0.01). The results showed that the lamellar cerebellum mainly experienced cell proliferation, differentiation and migration, and the lamellar cerebellum matured gradually. The death of the outer granular layer was mainly cell migration and apoptosis. The other layers mainly undergo cell differentiation. Development and apoptosis. P0 P14 is the period of rapid development of cerebellar cortical neurons, and also the peak period of apoptosis. The apoptotic pathway of Caspase-3 by activating the activation of Caspase-8 exists in cerebellar cortical plastic maturation.
【學位授予單位】：河南大學
【學位級別】：碩士
【學位授予年份】：2007
【分類號】：R33

【參考文獻】

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1 鄧錦波，蔡琰，，邱建勇，鞠躬，戴洪，孫曉江，王s

本文編號：1533856