| In vitro Assay for Dendritic Spine Retraction of Hippocampal Neurons with Sparse Labeling
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Author:
Date:
2016-09-20
[Abstract] Dendritic spines are the post-synaptic structures that play a central role in excitatory synaptic transmission. Developmental spinogenesis relies on a variety of stimuli such as those derived from cell-cell communication and their downstream signaling. Here, we describe an in vitro assay of dendritic spine retraction using hippocampal slice culture, in which individual neurons are sparsely and brightly labeled by the Supernova method, for the study of molecular mechanisms of spine development.
[摘要] 树突状棘是突触后结构，在兴奋性突触传播中起重要作用。 发育性的发生依赖于各种刺激物，如来自细胞 - 细胞通讯及其下游信号传导的刺激。 在这里，我们描述了使用海马片段培养的树突状脊柱退缩的体外测定，其中单个神经元被Supernova方法稀疏和明亮地标记，用于研究脊柱发育的分子机制。
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| In utero Electroporation of the Embryonic Mouse Retina
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Author:
Date:
2014-10-05
[Abstract] This protocol is useful to manipulate gene expression in the embryonic retina and compare the result with the contralateral non electroporated retina. In addition, the electroporation of a membrane or cytoplasmic tagged GFP allows to determine the effects of gene manipulation on the outgrowth of retinal ganglion cell axons (Garcia-Frigola et al., 2007) or simply to follow axon outgrowth in mutant embryos. DNA can be directed to different quadrants of the retina (ventral or dorsally) by modifying the position of the electrodes (Petros et al., 2009; Sánchez-Arrones et al., 2013). After the procedure, embryos are left developing to the desired stage, including postnatal stages.
[摘要] 该方案可用于操纵胚胎视网膜中的基因表达，并将结果与对侧非电穿孔视网膜进行比较。 此外，膜或细胞质标记的GFP的电穿孔允许确定基因操作对视网膜神经节细胞轴突的生长的影响（Garcia-Frigola等人，2007）或简单地遵循突变体胚胎中的轴突生长。 通过修改电极的位置，DNA可以被引导到视网膜的不同象限（腹侧或背侧）（Petros等，2009;Sánchez-Arrones等，2013）。 手术后，胚胎发育到期望的阶段，包括产后阶段。
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| In utero Electroporation of Mouse Embryo Brains
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Author:
Date:
2012-07-20
[Abstract] This is a non-invasive technique to introduce transgenes into developing brains. In this technique, DNA is injected into the lateral ventricle of the embryonic brains, and is incorporated into the cells through electroporation. Embryos then continue their development in normal conditions in vivo. The effects of genes of interest can be evaluated at certain time points after in utero electroporation. This technique allows acute knockdown or over expression of genes of interest. Compensatory effects from other genes are less likely to happen; it also circumvents possible chronic detrimental effects.
[摘要] 这是将转基因引入发育中的大脑的非侵入性技术。 在该技术中，将DNA注入胚胎脑的侧脑室，并通过电穿孔掺入细胞中。 胚胎然后在正常条件下在体内继续其发育。 可以在子宫内电穿孔后的某些时间点评估感兴趣的基因的效应。 这种技术允许急性敲减或过表达感兴趣的基因。 其他基因的补偿效应不太可能发生; 它也绕过可能的慢性有害影响。
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