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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Author:
Date:
2020-09-05
[Abstract] Human neuron transplantation offers novel opportunities for modeling human neurologic diseases and potentially replacement therapies. However, the complex structure of the human cerebral cortex, which is organized in six layers with tightly interconnected excitatory and inhibitory neuronal networks, presents significant challenges for in vivo transplantation techniques to obtain a balanced, functional and homeostatically stable neuronal network. Here, we present a protocol to introduce human induced pluripotent stem cell (hiPSC)-derived neural progenitors to rat brains. Using this approach, hiPSC-derived neurons structurally integrate into the rat forebrain, exhibit electrophysiological characteristics, including firing, excitatory and inhibitory synaptic activity, and establish ...
[摘要] [摘要] 人类神经元移植为建模人类神经系统疾病和潜在的替代疗法提供了新的机会。然而,人脑皮层的复杂结构分为六层,具有紧密互连的兴奋性和抑制性神经元网络,这对体内移植技术获得平衡,功能稳定和稳态稳定的神经元网络提出了重大挑战。在这里,我们提出了一项协议,将人类诱导的多能干细胞(hiPSC )衍生的神经祖细胞引入大鼠脑。使用这种方法,hiPSC 衍生的神经元在结构上整合到大鼠前脑中,表现出电生理特性,包括放电,兴奋性和抑制性突触活性,并与宿主电路建立神经元连通性。
[背景] 人类大脑皮层是一个复杂的细胞镶嵌体,在不同的皮质层(I-VI)中包含多样化的神经元亚型,可建立轴突输出和树突状输入的特定模式,提供了皮质电路的基本底物(Rakic,2009; Lodato 等等人,2011; Lui 等人,2011)。特别地,兴奋性和抑制性神经传递的平衡对于适当的脑功能是必需的(Turrigiano和Nelson,2004)。人类诱导的多能干细胞(hiPSC )可以在人类遗传背景下对人类神经系统疾病进行建模(Dolmetsch和Geschwind,2011; Brennand 等,2015; Vera和Studer,2015)。建立体外系统以将hiPSCs ...
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