| RNase Sensitivity Screening for Nuclear Bodies with RNA Scaffolds in Mammalian Cells
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Author:
Date:
2017-04-20
[Abstract] The mammalian cell nucleus is highly organized and contains membraneless nuclear bodies (NBs) characterized by distinct resident factors. The NBs are thought to serve as sites for biogenesis and storage of certain RNA and protein factors as well as assembly of ribonucleoprotein complexes. Some NBs are formed with architectural RNAs (arcRNAs) as their structural scaffolds and additional NBs likely remain unidentified in mammalian cells. Here, we describe an experimental protocol to search for new NBs built on certain arcRNAs. RNase-sensitive NBs were identified by monitoring nuclear foci visualized by tagging thousands of human cDNA products.
[摘要] 哺乳动物细胞核高度组织，包含以不同的居民因素为特征的无膜核体（NBs）。 NB被认为是用于某些RNA和蛋白质因子的生物发生和储存的位点以及核糖核蛋白复合物的组装。 一些NB由构建的RNA（arcRNA）形成，作为它们的结构性支架，另外的NB可能在哺乳动物细胞中保持不明。 在这里，我们描述了一个实验协议来搜索建立在某些arcRNA上的新NB。 通过监测通过标记数千个人类cDNA产物可视化的核病灶来鉴定RNase敏感性NB。
【背景】哺乳动物细胞核是高度组织的，由称为核体（NB）的多个不同的亚结构组成。迄今为止，已经将15个NB鉴定为含有各种蛋白质和RNA因子的亚核膜无颗粒结构，其中许多颗粒结构用作特异性RNA，蛋白质和/或核糖核蛋白（RNP）复合物的生物发生，成熟，储存和螯合的位点Mao et al。，2011; Sleeman and Trinkle-Mulcahy，2014）（表1）。
一些NB被构建在称为结构RNA（arcRNA）的特定长非编码RNA（lncRNA）上，其定义为NB的结构核心（Chujo等，2016）。 arcRNA依赖的NB由与arcRNA相互作用的许多RNA结合蛋白组成。最显着的例子是由几种特征性RNA结合蛋白组成的寄生虫斑（Fox等，2002; ...
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| Immunogold Labeling Analysis of Cell Wall Polysaccharides with Special Reference to (1;3,1;4)-β-D-glucan in Rice Cell Walls
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Author:
Date:
2016-03-05
[Abstract] Various types of cell wall compositions have evolved to fulfill a wide range of biological roles during the diversification of land plants. (1;3,1;4)-β-D-glucan (MLG) is a defining feature of the cell walls in the order Poales (Yokoyama and Nishitani, 2004), which has multiple functions associated with metabolic, growth, and defense systems. MLG is also a characteristic component of the matrix polysaccharides that undergo turnover and metabolism, depending on the tissue and the stage of development (Kido et al., 2015). Determining the extracellular localization of MLG is essential for elucidating its functions. Electron microscopy immunogold labeling analysis is a useful technique, which provides an accurate representation of the extracellular distribution of MLG. This strategy ...
[摘要] 各种类型的细胞壁组合物已经发展以在陆地植物的多样化过程中实现广泛的生物学作用。 （1; 3,1; 4）-β-D-葡聚糖（MLG）是Poales（Yokoyama和Nishitani，2004）中的细胞壁的定义特征，其具有与代谢，生长和防御相关的多种功能 系统。 MLG也是经历翻转和代谢的基质多糖的特征组分，这取决于组织和发育阶段（Kido等人，2015）。 确定MLG的细胞外定位对于阐明其功能是必要的。 电子显微镜免疫金标记分析是一种有用的技术，其提供MLG的细胞外分布的精确表示。 该策略也适用于各种细胞壁多糖，其在调节每种植物物种的生长和分化中具有关键作用。
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| Histochemical Staining of Silica Body in Rice Leaf Blades
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Author:
Date:
2015-10-05
[Abstract] Silicon (Si) is a biologically important element for plants in the order Poales (Yamamoto et al., 2011; Kido et al., 2015). In rice, Si is mainly deposited in the motor cells and the cell walls of the leaf epidermis. However, the molecular basis of this overall process has not been elucidated. Thus, we propose a protocol for the histochemical staining of the silica body based on specific hydrogen bonding between silanol group and the carboxylate group of crystal violet lactone (Ichimura et al., 2008), as described by Isa et al. (2010), but with minor modifications. This modified protocol can be used for observing Si accumulation during rice development.
[摘要] 硅（Si）对于Poales（Yamamoto等人，2011; Kido等人，2015年）的植物是重要的生物元素。 在稻中，Si主要沉积在运动细胞和叶表皮的细胞壁中。 然而，该整个方法的分子基础尚未阐明。 因此，我们提出了用于基于硅烷醇基团和结晶紫内酯的羧酸酯基团之间的特异性氢键的二氧化硅体的组织化学染色的方案（Ichimura等人，2008），如Isa （2010），但稍作修改。 这个修改的协议可以用于观察水稻开发过程中的硅积累。
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