| Optogenetic Mapping of Synaptic Connections in Mouse Brain Slices to Define the Functional Connectome of Identified Neuronal Populations
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Author:
Date:
2017-01-05
[Abstract] Functional connectivity in a neural circuit is determined by the strength, incidence, and neurotransmitter nature of its connections (Chuhma, 2015). Using optogenetics the functional synaptic connections between an identified population of neurons and defined postsynaptic target neurons may be measured systematically in order to determine the functional connectome of that identified population. Here we describe the experimental protocol used to investigate the excitatory functional connectome of ventral midbrain dopamine neurons, mediated by glutamate cotransmission (Mingote et al., 2015). Dopamine neurons are made light sensitive by injecting an adeno-associated virus (AAV) encoding channelrhodopsin (ChR2) into the ventral midbrain of DATIREScre mice. The efficacy and ...
[摘要] 神经回路中的功能连通性由其连接的强度，发病率和神经递质特性决定（Chuhma，2015）。使用光遗传学，可以系统地测量识别的神经元群体和定义的突触后靶神经元之间的功能性突触连接，以便确定该识别群体的功能性连接群。这里我们描述了用于研究由谷氨酸共转播介导的腹侧中脑多巴胺神经元的兴奋性功能性连接体的实验方案（Mingote等，2015）。通过将编码通道视紫红质（ChR2）的腺相关病毒（AAV）注射到DATIREScre小鼠的腹侧中脑中，使多巴胺神经元变得光敏。多巴胺合成酶酪氨酸羟化酶的免疫荧光证实ChR2表达在多巴胺神经元中的功效和特异性。然后，切片膜片钳记录由接受多巴胺神经元投影的区域中的神经元产生，并且确定兴奋性连接的发生率和强度。所有接受多巴胺神经元投射的区域的连接发生率和强度的总结构成功能性连接体。
【背景】为了建立特定神经回路的功能，有必要确定解剖连接，解剖连接的映射及其功能连接，连接的强度，发病率和神经递质性质的映射。使用单因素限制的病毒性突触后追踪技术，可以描述包括多巴胺系统在内的神经回路的复杂解剖连接（Callaway and Luo，2015; ...
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| Skeletal Myogenesis in vitro
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Author:
Date:
2015-11-05
[Abstract] Mature skeletal myofibers are elongated and multinucleated cells. Many stem/progenitor cell types, including committed muscle stem (satellite cells) and progenitor (myoblasts) cells, muscle-derived stem cells, myogenic endothelial cells, and mesenchymal stem/stromal cells, have been shown to exhibit skeletal myogenesis under appropriate inductive conditions. Committed muscle stem/progenitor cells and multipotent stem/progenitor cells which have skeletal myogenic capacity can typically be differentiated into skeletal myofibers in vitro following extended low-serum exposure. Differentiated cells exhibit distinct fiber-like elongated morphology with multiple nuclei and express unique muscle molecular markers indicating myogenesis, including desmin (early) and fast- and/or ...
[摘要] 成熟骨骼肌纤维是细长和多核细胞。 许多干/祖细胞类型，包括定型肌干（卫星细胞）和祖细胞（成肌细胞）细胞，肌肉衍生的干细胞，肌肉内皮细胞和间充质干/基质细胞，已显示在适当的诱导条件下表现出骨骼肌发生 。 具有骨骼肌生成能力的承诺的肌肉干/祖细胞和多能干/祖细胞可以在延长的低血清暴露后通常在体外分化成骨骼肌纤维。 分化的细胞表现出不同的纤维样细长形态与多个核，并表示独特的肌肉分子标记指示肌发生，包括结蛋白（早）和快和/或慢肌球蛋白重链（成熟）。
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