Wistar大鼠海马CA1区、CA3区和齿状回区的解剖分割

Division of CA1,CA3 and DG regions of the hippocampus of Wistar rat

目的:在体视显微镜下分割Wistar大鼠海马CA1区、CA3区和齿状回(DG)区。方法:24只健康Wistar大鼠,分组如下:①6只大鼠取脑后硫堇染色,观察海马各区细胞形态;②6只大鼠分离出海马,体视显微镜下观察海马形态并分割CA1区、CA3区和DG区,各区分别切片后硫堇染色;③12只大鼠检测海马各区HSP 70的表达。结果:①大脑冠状切片硫堇染色清晰显示出海马CA1区、CA3区和DG区;②体视显微镜下,在海马腹侧面,沿着CA1区和DG区之间的海马沟可分割开CA1区和DG区,沿着CA3区和DG区之间的裂隙可分割开CA3区和DG区;分割后的海马各区细胞形态结构与整体大脑冠状切片上相对应区域的细胞形态结构一致;③Western blot结果显示:与对照组相比,脑缺血组HSP 70的表达在海马CA3+DG区明显上调、而在CA1无明显变化,这一结果与免疫组织化学结果一致。结论:上述方法可比较明确地分割Wistar大鼠海马CA1区、CA3区和DG区,分割得到的各区组织可用于蛋白质表达的检测。

Objective： Division of the CA1,CA3 and dentate gyrus（DG） regions of the hippocampus of Wistar rat under the stereomicroscope.Methods： Twenty-four Wistar rats were randomly assigned to three groups.①The brain was sectioned coronally（n=6）.The sections were stained with thionin and the morphology of cells in each region of the hippocampus was observed under microscopy.②The hippocampus was dissected out and observed on the whole.Then,the CA1,CA3 and DG regions of the hippocampus were divided.Every region divided was sectioned,and the morphology of cells was observed.③Rats with brain ischemia or not were also decapitated and the HSP 70 expressions were observed in CA1,CA3 ＋ DG regions by Western blot and immunohistochemical staining（n=12）.Results： ①The CA1,CA3 and DG regions of the hippocampus could be clearly observed in coronal section of the brain stained by thionin.②Under the stereomicroscope,the CA1 and DG regions of the hippocampus could be separated along the hippocampal fissure between them in ventral surface of the hippocampus.The CA3 and DG regions of the hippocampus could be separated along a fissure between them.The appearance of cells in the sections of the divided CA1,CA3 and DG specimens is consistent with that in the brain coronal sections,respectively.③The results of Western blot indicated that the HSP 70 expression of the brain ischemia group was up-regulated significantly in CA3＋DG regions compared with the sham group.However,HSP 70 expression has no significant changes in CA1 region.The above results were consistent with those of the immunohistochemical staining.Conclusion： The CA1,CA3 and DG regions of the hippocampus of Wistar rat could be divided under stereomicroscope,and the divided each region was sensible for detection of protein using Western blot.