| 钟 磊,谢燕萍,杨 娟,冯伟伟,游 宇.小鼠海马神经元冰冻切片和脑片培养方法的比较[J].现代生物医学进展英文版,2017,17(28):5416-5420. |
| 小鼠海马神经元冰冻切片和脑片培养方法的比较 |
| Comparison of Methods for Frozen Section and Cultivate Slices of Hippocampal Pyramidal Neurons of Mouse |
| Received:April 02, 2017 Revised:April 29, 2017 |
| DOI：10.13241/j.cnki.pmb.2017.28.004 |
| 中文关键词: 冰冻切片 脑片培养 海马神经元 |
| 英文关键词: Frozen section Cultivate slices Pyramidal neurons |
| 基金项目:国家自然科学基金青年基金项目(81301116) |
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| 中文摘要: |
| 摘要 目的：通过冰冻切片和脑片培养方式比较获得更适合脑片实验研究的方法。方法：分别运用急性切片和脑片培养方法，结合全细胞膜片钳技术比较两种脑片处理方法对小鼠海马神经元细胞形态、细胞膜封接难易程度、细胞电生理特性等的差异，获得更适合细胞研究的脑片获取方法。结果：冰冻切片方法切断部分神经纤维，脑片表层出现肿胀或坏死细胞，2-3层细胞可用于膜片钳记录，但不易封接破膜。脑片培养后可使纤维再生，整个脑片细胞形态清晰可见，容易封接破膜，电生理记录波形及基本特性与冰冻切片一致，但脑片培养方法的细胞突触后电流幅度更大、频率更高。结论：脑片培养可修复受损纤维和细胞膜柔韧性，且不改变膜特性，但脑片培养重建了一定数量的细胞间信号连接，使细胞反应性增强，脑片培养方法更适合脑片神经元研究。 |
| 英文摘要: |
| ABSTRACT Objective: To obtain the more suitable method of brain slice experiment through comparing the methods of frozen sections and brain slice culture. Methods: Frozen section and brain slice culture were used respectively. The differences in mouse hippocampal neurons cell morphology, difficulty of cell membrane sealing degree, the electrophysiological properties of cells of two kinds of brain slice processing methods were compared using whole cell patch clamp technique to find out which way is more suitable for brain slice research. Results: Frozen section cut off part of the nerve fibers, leading to surface slices of cells swelling or necrosis. 2-3 layers of cells could be used for patch clamp recording, but it was not easy to break the sealing. Brain slice culture could make fiber regeneration. The cell morphology was clearly visible, and the membranes were easily ruptured. The electrophysiological recording waveform and the basic characteristics were consistent with those of frozen section, but brain slice culture has more larger on postsynaptic cell current amplitude and higher on frequency. Conclusion: Brain slice culture can repair the damaged cell membrane and fiber flexibility, and does not change the membrane properties. However, it can reconstruct a certain number of intercellular signal connection to enhance cell reactivity. Therefore, cultivate slices are more suitable for the study of neurons. |
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