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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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Author:
Date:
2020-05-05
[Abstract] The Ras homologous protein (Rho) GTPase subfamily, including RhoA, RhoB, and RhoC are small molecules (~21 kDa) that act as molecular switches in a wide range of signaling pathways to orchestrate biological processes associated with both physiological and tumorigenic cellular states. The Rho GTPases are crucial regulators of actin cytoskeleton rearrangements and FA dynamics and are required for effective cell migration and invasion, as well as cell cycle progression and apoptosis. The Rho GTPases activity is regulated by conformational switching between GTP-bound (active) and GDP-bound (inactive) states. This GTP/GDP cycling is tightly controlled by the guanine nucleotide exchange factors (GEFs), which function as activators by catalyzing the exchange of GDP for GTP and by the ...
[摘要] [摘要] 包括RhoA，RhoB和RhoC在内的Ras同源蛋白（Rho）GTPase亚家族是小分子（〜21 kDa ），在广泛的信号传导途径中充当分子开关，以协调与生理和致瘤细胞状态相关的生物学过程。该Rho GTP酶是至关重要的调节肌动蛋白细胞骨架重排和FA动态和所需要的有效细胞迁移和侵袭，以及细胞周期进程和凋亡，其Rho GTP酶的活动是监管下的构象之间切换GTP结合（有源）和GDP -- GTP / GDP循环受鸟嘌呤核苷酸交换因子（GEF）严格控制，鸟嘌呤核苷酸交换因子（GEF）通过催化GTP的GDP交换和GTPase激活蛋白（GAP）来激活，从而实现水解作用。导致Rho GTPase失活的GTP 的详细信息。在此，我们描述了执行RhoB G-LISA活化测定以检测体外GTP负载的RhoB的水平的详细协议。这是第一个专门用于测量RhoB活化的比色测定。该方法是通过改编RhoA G-LISA激活检测试剂盒（Cytoskeleton，Inc.）而开发的，可在不到3 小时的时间内精确测量RhoB活性，这种快速方法可广泛用于评估GTP负载的RhoB 的水平在任何种类的细胞模型中，要了解RhoB激活在生理过程，疾病，致癌转化中的作用或在高通量筛选中发现药物的作用。
[背景 ] 尽管Rho GTP酶的RhoA，RhoB的，和RhoC占有率超过85 ...
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