激活Rho激酶对大鼠海马神经元突起生长影响的原子力显微镜观察

发布时间：2018-06-23 18:12

本文选题：Rho激酶 + 海马神经元　；参考：《暨南大学》2008年硕士论文

【摘要】： 目的: 了解激活Rho激酶通路后,神经元突起的坍塌过程,细胞膜表面结构的变化,丰富突起形成的形态学资料。材料和方法: 体外原代培养大鼠海马神经元,用Rho激酶促进剂LPA干预海马神经元,原子力显微镜观察细胞突起的生长过程和细胞膜表面结构的变化趋势。结果: Neurobasal和B27无血清培养基培养获得的大鼠海马神经元纯度较高,且细胞生长活力旺盛,贴壁后6-11d的细胞适宜用于进行结构和功能等实验观察。随着神经元的发育,细胞突起不断生长,分支逐渐增多,其长度、直径和数量在观察的48小时达到高峰,之后略有下降。原子力显微镜显示出清晰的突起,神经元细胞膜结构丰富,主要包括膜蛋白、孔洞样结构等。在神经元发育的早期可见大量卵石样的膜蛋白嵌于细胞膜上,其表面凸凹不平,随着细胞发育的不断成熟,胞膜逐渐平整,而且出现较多的形状各异的孔洞样结构。用LPA干预后,神经元的突起明显较对照组缩短,而且其数量、长度、直径均变化明显,但是它们的变化趋势与对照组基本相似,均以48小时为主峰。细胞膜的变化主要表现在其不平整性维持相对较长的时间,有较多的膜蛋白富集,但在发育的后期仍出现孔洞样结构,其在数量和形状、大小均有别于对照组。结论: 神经元的发育是一个渐进过程,突起不断生长、延伸、分支逐渐增多,膜结构逐渐完善。Rho激酶活性提高可诱导突起坍塌,表现为突起缩短、数目减少。Rho激酶活性增加诱导膜结构发生改变,主要表现为物质转运结构的增多以及膜蛋白质丰富。
[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.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R329

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