对于高分辨率延时成像神经细胞迁移的一个器官型产后脑切片检测
Summary
本协议描述了一个器官的切片检测，产后脑和延髓迁徙流时间推移高分辨率成像成神经细胞迁移的优化。
Abstract
产后大脑中的神经再生取决于三个生物事件的保养：祖细胞，成神经细胞迁移，以及分化和整合到现有的神经回路的新的神经元的增殖。在嗅球产后神经发生，这些事件被隔离在三个不同的域解剖学：扩散主要是发生在侧脑室室管膜下区（SEZ），移植的神经母细胞遍历喙迁徙流（RMS），和新的神经细胞分化和内的嗅球（OB）的整合。这三个领域作为理想的平台来研究细胞，分子和生理机制调节的生物事件明显。本文介绍了一个器官产后脑组织切片检测进行了优化，其中外条件紧密地模仿迁移神经母细胞在体内环境。我们表明，我们的检测提供了统一，导向，迅速运动神经母细胞内的RMS。此法将是非常适合的细胞自主和非自主神经细胞迁移的调节，利用交叉移植的方法，从不同的遗传背景的小鼠的研究。
Protocol
一，程序下列技术应在无菌条件下进行，在层流罩，使用消毒工具。 器官片玻璃底部菜的制备菜肴必须在无菌环境下编写的，使用消毒工具。 一个片介质150μL下降（见食谱）被放置在玻璃底部的一部分菜小心，以避免气泡在中期中心。 随着一个一次性注射器与一个23计针，胶水多个点（橡皮泥 – 埃尔默的，猫＃E904）配备都放置在邻近?…
Discussion
在RMS的神经元的迁移，是产后神经发生在1嗅球的重要组成部分。通过RMS的迁移发生在大脑表面相切的平面。切向迁移的神经母细胞是从他们的祖源的位置，以及其神经元的最终产品1，2，3的发散命运的放射状迁移细胞的独特。切线方向迁移的细胞在产后RMS相对纯净的人口使这种解剖学上定义的区域研究切向迁移的机制的最佳平台。破译神经细胞迁移的细胞和分子机制的关键是理?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
我们感谢丹McWhorter在视频叙述议定书。这项工作是由美国国立卫生研究院批准5R01NS062182，衰老研究从美国联邦补助金，并授予HTG机构资金支持。
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Tags
Organotypic Slice AssayHigh-resolution Time-lapse ImagingNeuronal MigrationPostnatal BrainNeurogenesisProgenitor CellsSubependymal Zone (SEZ)Rostral Migratory Stream (RMS)Olfactory Bulbs (OB)Cellular MechanismsMolecular MechanismsPhysiological MechanismsExtracellular ConditionsMigrating NeuroblastsUniform MovementOriented MovementSpeedy MovementCell Autonomous RegulationNon-autonomous RegulationCross-transplantation Approaches
Cite This Article
Jacquet, B. V., Ruckart, P., Ghashghaei, H. T. An Organotypic Slice Assay for High-Resolution Time-Lapse Imaging of Neuronal Migration in the Postnatal Brain. J. Vis. Exp. (46), e2486, doi:10.3791/2486 (2010).