| Histochemical Staining of Suberin in Plant Roots
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Author:
Date:
2021-02-05
[Abstract] Histological stains are useful tools for characterizing cell shape, arrangement and the material they are made from. Stains can be used individually or simultaneously to mark different cell structures or polymers within the same cells, and to visualize them in different colors. Histological stains can be combined with genetically-encoded fluorescent proteins, which are useful for understanding of plant development. To visualize suberin lamellae by fluorescent microscopy, we improved a histological staining procedure with the dyes Fluorol Yellow 088 and aniline blue. In the complex plant organs such as roots, suberin lamellae are deposited deep within the root on the endodermal cell wall. Our procedure yields reliable and detailed images that can be used to determine the suberin pattern in ...
[摘要] [摘要]组织学染色是表征细胞形状,排列和其制成材料的有用工具。可以单独或同时使用污渍来标记同一细胞内的不同细胞结构或聚合物,并以不同的颜色对其进行可视化。组织学染色剂可以与遗传编码的荧光蛋白结合使用,这对于理解植物的发育非常有用。为了通过荧光显微镜观察木栓质片状细胞,我们改进了荧光黄088和苯胺蓝染料的组织学染色程序。在复杂的植物器官中,例如根部,木栓质薄片沉积在根部内的内胚层细胞壁上。我们的程序产生了可靠而详细的图像,可用于确定根细胞中的suberin模式。此协议的主要优点是它的效率,木栓质本地化的详细可视化它产生的根源,并返回到共焦图象进行分析和重新的可能性-评估数据如果必要的话。
[背景] Suberin是一种复杂的聚酯,与角质和木质素一起在陆地植物中形成物理屏障。在拟南芥(Arabidopsis thaliana )中,木栓质主要积聚在表皮叶细胞的细胞壁,种皮,周皮和根部内胚层细胞的细胞壁中。从根本上说,木栓质沉积是动态过程(Barberon等,2016; Andersen等,2018; Ursache等,2020),其受各种营养胁迫,干旱胁迫和植物激素信号传导途径的调节(Baxter等)。等人,2009; Kosma等人,2014; ...
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| Immuno-electrophysiology on Neuromuscular Junctions of Drosophila Third Instar Larva
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Author:
Date:
2021-02-05
[Abstract] Alterations in synaptic transmission are critical early events in neuromuscular disorders. However, reliable methodologies to analyze the functional organization of the neuromuscular synapses are still needed. This manuscript provides a detailed protocol to analyze the molecular assembly of the neuromuscular synapses through immune-electrophysiology in Drosophila melanogaster. This technique allows the quantification of the molecular behavior of the neuromuscular synapses by correlating the structural configuration of the synaptic boutons with their electrical activity.
[摘要] [摘要]突触传递的改变是神经肌肉疾病的关键早期事件。但是,仍然需要可靠的方法来分析神经肌肉突触的功能组织。此马努脚本提供了详细的协议,通过在免疫电分析神经肌肉突触的分子组装果蝇黑腹果蝇。该技术通过使突触钮扣的结构构型与其电活动相关联,可以量化神经肌肉突触的分子行为。
[背景]果蝇三龄幼虫的神经肌肉接头(NMJ)的功能组织在研究突触形成和功能的分子机制方面具有突出的优势(Feiguin等,2009 ; Godena等,2011; Romano等。(2014年和2015年;Strah等人,2020年),它在包括人类在内的其他物种中似乎是保守的。在这方面,果蝇NMJs的解剖组织是由多个突触钮扣构成的,这些突触钮扣是在运动轴突的最终乔木形成的。这些结构的分化和维持负责骨骼肌的神经支配,并暗示不同分子的协调作用,专门用于建立细胞接触以及释放,接收和整合神经递质信号传导所需的机制。此外,在果蝇中开发的功能强大的遗传工具以及神经系统对单个神经元的解剖学解析,提供了难得的机会,可以在可区分细胞的组织中进行全基因组范围的分子表型无偏搜索。尽管果蝇已经存在可视化神经肌肉突触建立的单独协议(Sabeva和Bykhovskaia,2017 ; Goel等人,2019 ...
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