| Laser Scanning Confocal Microcopy for Arabidopsis Epidermal, Mesophyll, and Vascular Parenchyma Cells
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Author:
Date:
2017-03-05
[Abstract] Investigation of protein targeting to plastids in plants by confocal laser scanning microscopy (CLSM) can be complicated by numerous sources of artifact, ranging from misinterpretations from in vivo protein over-expression, false fluorescence in cells under stress, and organellar mis-identification. Our studies have focused on the plant-specific gene MSH1, which encodes a dual targeting protein that is regulated in its expression and resides within the nucleoid of a specialized plastid type (Virdi et al., 2016). Therefore, our methods have been optimized to study protein dual targeting to mitochondria and plastids, spatial and temporal regulation of protein expression, and sub-organellar localization, producing a protocol and set of experimental standards that ...
[摘要] 通过共焦激光扫描显微镜（CLSM）对植物中质体进行蛋白质靶向的研究可能由于许多来源的伪像而复杂化，其范围从体内蛋白质过度表达的误解，应激细胞中的假荧光，和细胞器错误识别。我们的研究集中在植物特异性基因MSH1上，其编码双重靶向蛋白，其在其表达中被调节并且位于特定质体类型的核内（Virdi等人，2016）。因此，我们的方法已被优化，以研究蛋白质双重靶向线粒体和质体，蛋白质表达的空间和时间调节和亚细胞定位，产生其他人可能对这些研究有用的方案和一组实验标准。
背景 植物中的蛋白质靶向行为受氨基末端前序列以及可影响亚组织定位行为的内部序列特征的影响（Baginsky和Gruissem，2004）。结合启动子驱动的表达空间和时间调节，蛋白质的活性可以通过时间和位置而非常精确和专一。在MSH1的情况下，这种核编码的植物特异性蛋白质是双重靶向线粒体和质体（Xu et al。，2011）。启动子特征将其表达指导到生殖，表皮和血管薄壁细胞（Virdi等人，2016）。内部蛋白质特征将其定位于线粒体和质体核，以及质体类囊体膜。使用这里描述的方法，通过激光扫描共聚焦显微镜大大促进了这些不寻常的蛋白质特征的发现。使用更传统的细分器分割方法，大部分细节将被忽略。
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| Dissection of Different Areas from Mouse Hippocampus
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Author:
Date:
2013-11-05
[Abstract] The hippocampus modulates a number of modules including memory consolidation, spatial navigation, temporal processing and emotion. A banana-shaped structure, the hippocampus is constituted of morphologically distinct subregions including the dentate gyrus, CA3 and CA1 (here, we do not distinguish the “hippocampus proper” which consists only of CA1, CA3 and smaller CA2 and CA4 areas, from the “hippocampal formation,” composed of these in addition to the dentate gyrus and subiculum). Distinct cell types give rise to unique axonal fiber pathways in the dentate gyrus, CA3 and CA1 subregions; accordingly, these areas may exhibit differential molecular profiles in response to a number of behavioral paradigms and pharmacological and genetic treatments. It is therefore in the interest of the ...
[摘要] 海马调节了许多模块，包括记忆整合，空间导航，时间处理和情感。 香蕉形结构，海马由形态上不同的子区域组成，包括齿状回，CA3和CA1（这里，我们不区分仅由CA1，CA3和较小的CA2和CA4区域组成的“海马属”， “海马形成”，除了齿状回和子宫外还包括这些）。 不同的细胞类型在齿状回，CA3和CA1亚区域产生独特的轴突纤维途径; 因此，这些区域可以响应于许多行为范例和药理学和遗传处理而呈现差异分子谱。 因此，调查人员有兴趣从整个海马剖析一个特定的次区域。 在这里，我们概述了一个关于成年小鼠子区域特异性解剖的方案。
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