目的 通过对不同reelin基因型小鼠海马区组织学结构的比较分析，为reelin在海马结构形成中的作用提供科学依据；方法 选择杂合子reelin小鼠繁殖后代，在出生后10天进行基因型鉴定，设reelin+/+、reelin+/-和reelin-/-三组，每组3只小鼠。鉴定后的小鼠通过4%多聚甲醛心脏灌注后剥离全脑，通过多聚甲醛固定、蔗糖脱水后冰冻切片，采用DAPI染色细胞核，用荧光免疫组化标记Tbr-1、NeuN和GFAP特异性表达的细胞，显微镜下观察拍照后进行结果的分析；结果 从DAPI染色的细胞核及Tbr-1、NeuN和GFAP标记的细胞结构可以看出，reelin+/+型小鼠和reelin+/-型小鼠海马区都具有相似的结构，但将reelin+/+和reelin-/-型小鼠相比较，reelin-/-型小鼠海马结构模糊，齿状回细胞向门区扩散，海马本部的细胞层结构发生紊乱，出现密集双细胞带，GFAP标记细胞减少；结论 Reelin的缺失表达对小鼠大脑正常结构的形成具有明显的影响，在reelin表达缺失后海马区齿状回细胞分布弥散，海马本部细胞层的分布也发生紊乱，胶质细胞及纤维分布减少。
Abstract
Objective To provide scientific basis data for the function of Reelin in the hippocampal formation by comparing the histological structure of the hippocampus in different reelin genotype mice. Methods Heterozygous reelin mice offspring were used, and genotype identification was performed after birth for 10 days. Experiment was divided into three groups including reelin+/+, reelin+/- and reelin-/-, and each group had 3 mice. After identification, the mice were perfused through heart by 4% paraformaldehyde and the whole brain wascollected. All samples were fixed by paraformaldehyde, sucrose dehydration and finally cut into frozen coronal slice. DAPI was used to stain nuclei, and fluorescence immunohistochemistry was performed to label Tbr-1, NeuN and GFAP specific expression cells, then fluorescence microscope was used for further observation and analysis; Results From the DAPI stained nuclei and Tbr-1, NeuN and GFAP labeled cell structure, it could be seen that reelin+/+ mice and reelin+/- mice in hippocampal area had similar structure, but in comparison with the reelin+/+ and reelin-/- mice, dentate gyrus cells of the reelin-/- type mouse spreaded to the gate area, the cell layer of hippocampal structure disordered, and GFAP labeled cells reduced; Conclusion Reelin has obvious effect on normal formation of mouse brain structure. Under the condition of reelin knock out mouse, the dentate gyrus area cells are dispersed, the cell layer of the hippocampus is disordered, and glial cells and fiber distribution both are reduced.
关键词
Reelin /
reeler小鼠 /
海马 /
组织学结构
Key words
Reelin /
reeler mice /
hippocampus /
histological structure
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