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  本文選題：Rho激酶 + 海馬神經(jīng)元��； 參考：《暨南大學(xué)》2008年碩士論文

【摘要】： 目的: 了解激活Rho激酶通路后,神經(jīng)元突起的坍塌過程,細胞膜表面結(jié)構(gòu)的變化,豐富突起形成的形態(tài)學(xué)資料。 材料和方法: 體外原代培養(yǎng)大鼠海馬神經(jīng)元,用Rho激酶促進劑LPA干預(yù)海馬神經(jīng)元,原子力顯微鏡觀察細胞突起的生長過程和細胞膜表面結(jié)構(gòu)的變化趨勢。 結(jié)果: Neurobasal和B27無血清培養(yǎng)基培養(yǎng)獲得的大鼠海馬神經(jīng)元純度較高,且細胞生長活力旺盛,貼壁后6-11d的細胞適宜用于進行結(jié)構(gòu)和功能等實驗觀察。 隨著神經(jīng)元的發(fā)育,細胞突起不斷生長,分支逐漸增多,其長度、直徑和數(shù)量在觀察的48小時達到高峰,之后略有下降。原子力顯微鏡顯示出清晰的突起,神經(jīng)元細胞膜結(jié)構(gòu)豐富,主要包括膜蛋白、孔洞樣結(jié)構(gòu)等。在神經(jīng)元發(fā)育的早期可見大量卵石樣的膜蛋白嵌于細胞膜上,其表面凸凹不平,隨著細胞發(fā)育的不斷成熟,胞膜逐漸平整,而且出現(xiàn)較多的形狀各異的孔洞樣結(jié)構(gòu)。 用LPA干預(yù)后,神經(jīng)元的突起明顯較對照組縮短,而且其數(shù)量、長度、直徑均變化明顯,但是它們的變化趨勢與對照組基本相似,均以48小時為主峰。細胞膜的變化主要表現(xiàn)在其不平整性維持相對較長的時間,有較多的膜蛋白富集,但在發(fā)育的后期仍出現(xiàn)孔洞樣結(jié)構(gòu),其在數(shù)量和形狀、大小均有別于對照組。 結(jié)論: 神經(jīng)元的發(fā)育是一個漸進過程,突起不斷生長、延伸、分支逐漸增多,膜結(jié)構(gòu)逐漸完善。Rho激酶活性提高可誘導(dǎo)突起坍塌,表現(xiàn)為突起縮短、數(shù)目減少。Rho激酶活性增加誘導(dǎo)膜結(jié)構(gòu)發(fā)生改變,主要表現(xiàn)為物質(zhì)轉(zhuǎn)運結(jié)構(gòu)的增多以及膜蛋白質(zhì)豐富。
[Abstract]:Aim: to understand the collapse process of neuronal processes and the changes of cell membrane surface structure after activation of Rho kinase pathway, and to enrich the morphological data of neurite formation. Materials and methods: primary cultured rat hippocampal neurons were treated with Rho kinase promoter LPA. The process of neurite growth and the change trend of cell membrane surface structure were observed by atomic force microscope (AFM). Results: the hippocampal neurons cultured in Neurobasal and B27 serum-free medium were of high purity and high cell growth activity. The cells of 6-11 days after adherent were suitable for structural and functional observation. With the development of neurons, the neurites grew and the branches increased. The length, diameter and number of the neurons reached the peak at 48 hours after observation, and then decreased slightly. Atomic force microscopy (AFM) showed clear protrusions and abundant cellular membrane structures, including membrane proteins and voids. In the early stage of neuronal development, a large number of pebble like membrane proteins were embedded in the cell membrane, and the surface of the membrane was uneven. As the cell matured, the cell membrane was gradually smooth, and there were more vore-like structures of different shapes. After LPA treatment, the neurites were shorter than those of the control group, and the number, length and diameter of the neurons were obviously changed, but the trend of the changes was similar to that of the control group, with 48 hours as the main peak. The change of cell membrane was mainly due to the relatively long time of its unevenness and the enrichment of membrane protein, but at the late stage of development, the pore structure was still present, and its quantity, shape and size were different from those of the control group. Conclusion: the development of neurons is a gradual process, with the process growing, extending, branching increasing, and the membrane structure gradually improving. The activity of Rho kinase can induce the collapse of the neurite, which is characterized by the shortening of the process. The increase of Rho kinase activity induced the change of membrane structure, which was mainly due to the increase of substance transport structure and the abundance of membrane protein.
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號】：R329

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相關(guān)期刊論文 前3條

1 宮偉;羅卓荊;韓驊;秦鴻雁;褚尤彪;胡學(xué)昱;蘭麗鋒;;體外分離培養(yǎng)Balb/c小鼠胚胎干細胞向神經(jīng)細胞定向誘導(dǎo)分化[J];第四軍醫(yī)大學(xué)學(xué)報;2006年10期

2 林玲;鄭志z，

本文編號：2057978