Rho激酶对大鼠海马神经元突起生长和细胞骨架分布的调节

发布时间：2018-04-18 09:01

本文选题：Rho激酶 + 突起生长　；参考：《暨南大学》2008年硕士论文

【摘要】： 目的:探讨Rho激酶对大鼠海马神经元突起生长和细胞骨架的调节。材料和方法:以原代大鼠海马神经元为观察对象,用Rho激酶激动剂LPA和抑制剂Y-27632改变细胞内Rho激酶的活性,分别用时差显微成像技术观察神经元突起生长的动态变化,激光共聚焦显微镜观察细胞骨架分布的变化。结果:(1)时差显微成像结果:对照组神经元一级、二级突起不断形成,向外延伸,在末端形成三级突起;LPA组神经元大部分突起进行性塌陷,突起数量减少,且较细小;Y-27632组神经元胞体上多个一级突起形成,延伸较远,再分支现象明显,在远侧二级、三级突起生长迅速;Y-27632+LPA组神经元一些细小的一级突起退缩消失,其余突起不断分支、延伸,一级、二级、三级突起数量和长度较LPA组增加;LPA+Y-27632组神经元一级突起逐渐停止塌陷,然后向外延伸,二级突起生长迅速,在其远侧可见三级突起和细小的四级突起形成。(2)激光共聚焦显微镜结果:神经元胞体、突起均有清晰的微管和微丝分布,在细小突起、突起末梢及丝状足内微丝分布较多;LPA组神经元与对照组相比,各级突起尤其是次级突起缩短,数量减少,微管、微丝在突起内分布减少明显,丝状足很少;Y-27632组神经元突起分支长且多,微管、微丝沿胞体和突起分布,突起远端及小分支上微丝的分布较多,有多个细长的丝状足在突起上形成;在LPA+Y-27632和Y-27632+LPA组神经元各级突起均较丰富,与LPA组相比微管和微丝表达有明显改善,在突起远端有较丰富的丝状足。结论:(1)抑制Rho激酶可促使神经元突起各级分支数目增加和突起生长。(2)激活Rho激酶可诱导神经元突起坍塌,表现为各级突起数目的减少和长度的缩短。(3)激活Rho激酶后再抑制或抑制Rho激酶后再激活,Rho激酶诱导神经元突起回缩的作用均下调,神经元突起继续生长,各级分支数目增加。(4)Rho激酶可以通过调节细胞骨架的分布参与突起的生长、延伸及分支形成。
[Abstract]:Objective: To investigate the regulation of Rho kinase on the growth and cytoskeleton of hippocampal neurons in rats.
Materials and methods: in cultured rat hippocampal neurons were observed, with Rho kinase inhibitor Y-27632 agonist LPA and the change of intracellular Rho kinase activity, respectively with the dynamic changes of neuronal growth observed time difference micro imaging technology, to observe the distribution of cytoskeleton in laser scanning confocal microscope.
Results: (1) the time difference imaging results: in the control group, neuron level, two level projections formed continuously extends outward to form three level projections at the end; LPA group of neurons most neurites collapsed, reducing the number of projections, and relatively small; forming a plurality of first level neurites in group Y-27632 neurons, extended a well, then the branch phenomenon is obvious in the far side of the two level, three level neurites grew rapidly; Y-27632+LPA group of neurons small first level neurites retraction disappeared, other neurites continued to branch, extension, level, level two, level three. The number and length were higher than that in LPA group; LPA+Y-27632 group of neurons level processes gradually stop collapse, and then extended outward, two level neurites grew rapidly and formed three small visible protrusions and the four protrusions on its far side. (2) by laser confocal microscopy: neurons, projections are microtubule and microfilament distribution in clear, fine process Since more microfilaments neurite endings and filopodia; neurons in the LPA group compared with the control group, especially secondary ecphyma neurites shortened, reduce the number of microtubules, microfilaments in neurites decreased significantly, filopodia rarely; group Y-27632 neurite branch length and number, microtubule, microfilament distribution along the cell bodies and processes. The distal projections and small branches of microfilament distribution and a plurality of elongated filopodia formation in neurites; in LPA+Y-27632 and Y-27632+LPA groups of neurons neurites were rich, compared with the LPA group the expression of microfilaments and microtubules had improved significantly, there is abundant in the distal neurite filopodia.
Conclusion: (1) inhibition of Rho kinase could promote neurite neurite growth and increase the number of branches. (2) activation of Rho kinase could induce neurite collapse, showed a reduction in the number of neurites and the length decrease. (3) after the activation of Rho kinase inhibition or inhibition of Rho kinase and Rho kinase activation. Induced neurites retraction effects were reduced, the neurites continued to grow, increasing the number of branches. (4) Rho kinase can regulate the cytoskeletal distribution involved in neurite outgrowth, extension and branching.

【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

【共引文献】