原代培养脊髓神经元神经丝蛋白NF-H表达与免疫电镜观察

Study on Expression of Neurofilament-H(NF-H) in Primary Cultured Neurons of Spinal Cord and Its Distribution under Immuno-electron Microscope

目的:研究原代培养脊髓神经元NF-H表达及免疫电镜的分布,为神经丝结构异常所致的神经疾病奠定实验基础。方法:采用神经细胞培养技术、免疫组织化学与免疫电镜技术,进行原代培养脊髓神经元的神经丝蛋白NF-H的表达,并用图像分析仪对NF-H表达进行了定量分析测定。结果:原代培养脊髓神经元中,神经丝蛋白的NF-H的表达,随细胞发育的逐渐成熟呈日趋上升趋势。免疫电镜观察可见神经丝蛋白呈电子密度高的丝絮状沉淀,均匀分布在细胞质中,在突起中呈束状排列。结论:神经丝蛋白的NF-H表达的增加,伴随着神经元的发育初期到成熟的形态结构变化,与培养时间成正相关。

Objective:To study the expression of neurofilament-H( NF-H) in primary cultured neurons of spinal cord and the distribution under electron microscope demonstrated by immuno-EM method in order to provide the experimental basis for further research about the diseases caused by abnormal development of neurofilament. Methods; Cell culture, immunohistochemical and immuno-EM techniques were employed to investigate the expression of NF-H in primary cultured neurons of spinal cord and its distribution under the EM. Then semi-quantitative statistics analysis using microscopic Image-Analysis technique was conducted. Results; The expression of NF-H in primary cultured neurons of spinal cord increases correspondingly companied by cultured days. Under electron microscope, the NF-H appears as electron-dense thread-liked structures, which is well-distributed in soma and arranged in bundles in processes. Conclusion: The increasing of expression of NF-H in primary cultured neurons of spinal cord corresponds to morphologic changing of neurons during their developmental processes from immature to mature, and is closely related to cultured days.