Imaging activation of adult-generated granule cells in spatial memory
Imaging activation of adult-generated granule cells in spatial memory
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DOI：
10.1038/nprot.2007.415
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Frankland, Paul W.
中科院分区：
文献类型：
--
作者：
Kee, Nohjin;Teixeira, Catia M.;Frankland, Paul W.
在整个成年期，新的神经元在海马体的颗粒下区域不断地产生，人们对这些新的神经元是否在功能上整合到记忆回路中越来越感兴趣。这项协议描述了免疫组织化学程序，以可视化的新神经元招募到支持完整的小鼠的空间记忆回路。为了标记成人生成的颗粒细胞，小鼠被注射了增殖标记5-溴-2‘-脱氧尿苷(BrdU)。在BrdU治疗后的不同延迟，小鼠被训练在Morris水迷宫中找到一个隐藏的平台，然后可以在将平台从池中移除的探测测试中测试空间记忆。在探针测试90分钟后，对小鼠进行灌流，并对组织进行切片。免疫组织化学方法被用来定量BrdU标记的细胞和即刻早期基因Fos的表达。由于Fos的表达受神经元活动的调节，BrdU标记的神经元和Fos标记的神经元之间的重叠程度提供了一个迹象，表明成年产生的颗粒神经元是否已经整合到空间记忆回路中。
New neurons are continuously generated in the subgranular zone of the hippocampus throughout adulthood, and there is increasing interest as to whether these new neurons become functionally integrated into memory circuits. This protocol describes the immunohistochemical procedures to visualize the recruitment of new neurons into circuits supporting spatial memory in intact mice. To label adult-generated granule cells, mice are injected with the proliferation marker 5-bromo-2'-deoxyuridine (BrdU). At different delays after BrdU treatment, mice are trained to locate a hidden platform in the Morris water maze, and spatial memory can then be tested in a probe test with the platform removed from the pool. Ninety minutes after this probe test, mice are perfused and tissue is sectioned. Immunohistochemical procedures are used to quantify BrdU-labeled cells and expression of the immediate early gene, Fos. Because Fos expression is regulated by neuronal activity, the degree of overlap between BrdU-labeled and Fos-labeled neurons provides an indication of whether adult-generated granule neurons have been incorporated into spatial memory circuits.