| Optimized Immunostaining of Embryonic and Early Postnatal Mouse Brain Sections
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Author:
Date:
2021-01-05
[Abstract] The mammalian neocortex, the outer layer of the cerebrum and most recently evolved brain region, is characterized by its unique areal and laminar organization. Distinct cortical layers and areas can be identified by the protein expression of graded transcription factors and molecular determinants that define the identity of different projection neurons. Thus, specific detection and visualization of protein expression is crucial for assessing the identity of neocortical neurons and, more broadly, for understanding early and late developmental mechanisms and function of this complex system. Several immunostaining/immunofluorescence methods exist to detect protein expression. Published protocols vary with regard to subtle details, which may impact the final outcome of the immunofluorescence. ...
[摘要] [摘要]哺乳动物的新皮层是大脑的外层，也是最近进化的大脑区域，其特征在于其独特的区域和层状组织。不同的皮质层和区域可以通过分级转录因子的蛋白质表达和定义不同投射神经元身份的分子决定簇来鉴定。因此，蛋白质表达的特异性检测和可视化对于评估新皮层神经元的身份至关重要，更广泛地来说，对于理解这个复杂系统的早期和晚期发育机制和功能至关重要。一些 存在免疫染色/免疫荧光方法来检测蛋白质表达。已发布的方案在细节方面有所不同，可能会影响免疫荧光的最终结果。在这里，我们提供了详细的协议，适用于低温恒温器薄切片和厚振动切片器薄切片，该协议已成功地开发了针对新皮质发育关键分子的抗体。从早期的技术措施测距的大脑采集到的图像分析和统计，我们包括有关样品包容和每一个细节部分ING，幻灯片存储和旨在减少非特异性背景最佳抗体稀释。在实验室中常规使用的，我们的背景优化的免疫染色协议允许区域的有效检测-和层-独特新皮层投射神经元的特定的分子决定因素。
图形摘要：
优化的小鼠脑切片免疫染色方案的工作流程图。一。一个流程图的不同步骤的优化免疫染色协议薄低温恒温器和厚振动切片。乙。例如，用于在薄冠状免疫染色对SATB2和CTIP2部（20 μ ...
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| Peripheral Nerve Injury: a Mouse Model of Neuropathic Pain
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Author:
Date:
2017-05-05
[Abstract] Neuropathic pain is one of the highly debilitating chronic pain conditions, for which, currently, there is no therapeutic treatment. In order to reveal the underlying mechanism for neuropathic pain, various animal models have been established (Burma et al., 2016). This protocol describes how to prepare spinal nerve injury model (Kim and Chung, 1992; Rigaud et al., 2008; Masuda et al., 2016), one of the most frequently-used and highly reproducible models in which multiple alterations occur both in the peripheral and central nervous system.
[摘要] 神经性疼痛是高度虚弱的慢性疼痛病症之一，目前尚无治疗。为了揭示神经性疼痛的潜在机制，已经建立了各种动物模型（缅甸等人，2016）。该方案描述了如何准备脊髓神经损伤模型（Kim和Chung，1992; Rigaud等人，2008; Masuda等人，2016），其中最大经常使用和高度可重现的模型，其中在外周和中枢神经系统中发生多次改变。
背景 揭示神经性疼痛的潜在机制是开发有效治疗以达到最佳治疗的必要条件。因此，已经开发了用于神经性疼痛的各种动物模型。特别地，啮齿动物模型经常被使用，因为它们是高度可重复性的，并且在神经性疼痛患者中也观察到疼痛超敏反应。在本协议中，我们描述如何准备小鼠脊髓神经损伤模型。
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| Olfactory Bulb (OB) Transplants
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Author:
Date:
2016-09-05
[Abstract] Transplantation in mouse brain slices is a powerful tool in order to study axon targeting and migrational events during development. Taking advantage of donors and recipients belonging to different genotypes, this technique allows researchers to assess the contribution of donor and/or recipient tissue by performing various combinations and to study cell-autonomous functions or effects that are influenced by the recipient’s environment (Bastakis, et al., 2015). Here we describe the transplantation procedure on sagittal brain slices containing olfactory bulb (OB). Specifically, we have transplanted the proximal-to-the-cortex part of dorsal OB to the same region on a recipient slice. Transplanted slices can be cultured for up to 3 days before their morphology is disfigured due to ...
[摘要] 小鼠脑切片中的移植是一个强大的工具，以便研究轴突靶向和发展过程中的迁移事件。 利用属于不同基因型的供体和受体，该技术允许研究人员通过进行各种组合来评估供体和/或受体组织的贡献，并研究受受体环境影响的细胞自主功能或效应（Bastakis， em>等人，2015）。 在这里我们描述的矢状大脑切片包含嗅球（OB）的移植程序。 具体来说，我们已经将背侧OB的近端到外皮部分移植到受体切片上的相同区域。 移植的切片可以培养长达3天，之后由于3D中的生长它们的形态被破坏。 这些切片的重新切片允许更详细的免疫组织化学分析。
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