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应用免疫组化和组织化学方法显示学习记忆障碍小鼠脑中NOS神经元的变化.健康昆明小鼠32只,随机分为实验组和对照组,在立体定位仪上,以前国囟3mm,中线右旁开1.5mm,深1.5mm为座标,实验组向海马内注入acid0.5μl对照组海马内注入无菌生理盐水0.5μl.学习记忆测试,采用辐射式三等份Y型迷宫测试装置.术后7、14天小鼠经主动脉插管用4%多聚甲醛缓冲液灌注固定30分钟,-18℃下行脑连续冰冻切片,分别作NADPH-d组织化学反应和nNOS免疫组化反应.结果:(1)实验组小鼠术后Y型迷宫测试正确次数较术前明显减少,与对照组同时间点相比,差异有显著性,说明一侧海马注入KA使动物的空间辨别学习记忆能力受损.但术后14天实验组小鼠在迷宫测试中的正确次数渐有增加,表明其学习记忆能力有所恢复.(2)组织切片中NADPH-d组化染色和nNOS免疫化染色均显示皮质和海马内的NOS神经元.术后7、14天实验组与对照组,额、顶、颞叶皮质内的NOS神经元数目差异无显著性.在实验组,梨状皮质内的NOS神经元数目显示增多,胞体较小,主要分布于外颗粒层和锥体细胞层,形成一明显的带状;海马内NOS神经元主要分布于CAI-2区,CA3-4区较少,小鼠海马CAl-4区内的NOS神经元明显减少.nNOS免疫组化显示齿状四颗粒细胞层内NOS神经元数量较多,但这些神经元在NADPH-
Immunohistochemical and histochemical methods were used to show the changes of NOS neurons in the brain of learning and memory deficient mice.All 32 healthy Kunming mice were randomly divided into experimental group and control group.At stereotaxic instrument, The right side of open 1.5mm, 1.5mm deep as the coordinates, the experimental group injected into the hippocampus acid0.5μl control group into the hippocampus sterile saline 0.5μl learning and memory test using the radiation three equal parts of the Y-maze test device. At 7 and 14 days after operation, the mice were perfused with 4% paraformaldehyde buffer for a period of 30 minutes via aortic cannulation. Continuous brain sections were frozen at -18 ℃ for NADPH-d histochemistry and nNOS immunohistochemistry respectively. Results: (1) The number of postoperative Y-maze test in experimental group was significantly decreased compared with that before operation, which was significantly different from that in control group at the same time point However, the correct number of mice in the experimental group increased gradually in the maze test 14 days after operation, indicating that their learning and memory abilities have been restored. (2) NADPH-d histochemical staining and nNOS immunostaining Both NOS neurons in the cortex and hippocampus were shown There was no significant difference in the number of NOS neurons between frontal, parietal and temporal cortex in the experimental group and the control group.In the experimental group, the number of NOS neurons in the piriform cortex increased and the cell bodies were smaller, mainly distributed in the outer granular layer And the pyramidal cell layer, forming a distinct ribbon; NOS neurons in hippocampus were mainly distributed in CAI-2 region, CA3-4 region less, NOS neurons in hippocampal CAl-4 area of mice significantly reduced.nnOS immunization The number of NOS neurons in the layer of dentate quadrilateral granulosa cells was higher than that of the group of NADPH-