| Dissection and Whole Mount Staining of Retina from Neonatal Mice
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Author:
Date:
2018-10-05
[Abstract] Here we provide a detailed protocol for whole mount staining of mouse retina. This protocol was used to analyze retinal angiogenesis in newborn mice (Sawaguchi et al., 2017) by modifying the original protocols (Powner et al., 2012; Tual-Chalot et al., 2013). This protocol can also be used for whole mount staining of adult retina.
[摘要] 在这里，我们提供了小鼠视网膜整体染色的详细方案。 该方案用于分析新生小鼠的视网膜血管生成（Sawaguchi et al。，2017），修改原始方案（Powner et al。，2012; Tual-Chalot et al。，2013）。 该方案也可用于成人视网膜的整体染色。
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| Determination of H+-ATPase Activity in Arabidopsis Guard Cell Protoplasts through H+-pumping Measurement and H+-ATPase Quantification
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Author:
Date:
2017-12-20
[Abstract] The opening of stomata in plants in response to blue light is driven by the plasma membrane H+-ATPase in guard cells. To evaluate the activation of the H+-ATPase in vivo, we can use H+-pumping by guard cells in response to blue light and fusicoccin. To do this, it is required to prepare a large amount of guard cell protoplasts and measure H+-pumping in the protoplasts. It is also necessary to determine the protein amount of H+-ATPase. In this protocol, we describe the procedures required for these preparations and measurements.
[摘要] 响应蓝光的植物气孔的开放是由保卫细胞中的质膜H + -ATPase驱动的。 为了评价体内H + -ATP酶的激活，我们可以使用H + +保卫细胞对蓝光的响应，fusicoccin。 为此，需要制备大量的保卫细胞原生质体，并测量原生质体中的H + - 抽吸。 还需要确定H + -ATP酶的蛋白质量。 在这个协议中，我们描述了这些准备和测量所需的程序。
【背景】响应于蓝光的气孔的开放是由穿过保卫细胞质膜上的H +介导的膜超极化驱动的（Assmann等，1985; Shimazaki等人，1986），并且是由质膜H + -ATP酶引起的（Kinoshita和Shimazaki，1999）。 H + -ATP酶在膜上产生电化学梯度，并提供植物细胞中许多次级运输所需的能量。然而，测量体内H + -ATP酶活性并不容易。利用保卫细胞的蓝光敏感特性，我们的方法可以将体内H +泵送作为体内测量H + + 使用拟南芥保卫细胞原生质体的ATP酶活性（Ueno等人，2005）。与通过蛋白质印迹（Yamauchi等人，2016）的Hβ+ -ATPase定量一起，该方法允许比较Hβ+ -ATPase活性不同的条件或突变背景。
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| Purification of Flagellin from Acidovorax avenae and Analysis of Plant Immune Responses Induced by the Purified Flagellin
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Author:
Date:
2016-08-20
[Abstract] Plants sense potential pathogens by recognizing conserved pathogen-associated molecular patterns (PAMPs) that cause PAMP-triggered immunity (PTI) including the generation of reactive oxygen species, callose deposition, and expression of several PTI-related genes. Acidovorax avenae is a Gam-negative bacterium that causes a seedling disease characterized by the deposition of brown stripes on the sheaths of infected plants. We previously reported that flagellin isolated from the rice avirulent A. avenae N1141 strain induces PTI, while flagellin isolated from the rice virulent A. avenae K1 strain does not induce PTI. To examine the molecular mechanism of specific PTI induction by N1141 flagellin, highly purified flagellin from N1141 or K1 strains is required. Here, ...
[摘要] 植物通过识别引起PAMP触发的免疫（PTI）的保守的病原体相关分子模式（PAMP）来感测潜在的病原体，包括活性氧的产生，胼lose质沉积和几个PTI相关基因的表达。 酸性弧菌是一种革兰氏阴性细菌，其导致幼苗疾病，其特征在于在感染的植物的鞘上沉积棕色条纹。 我们以前报告了从无毒力的大米中分离的鞭毛蛋白。 avenae N1141菌株诱导PTI，而从水稻毒力分离的鞭毛蛋白。 avenae K1株不诱导PTI。 为了检查N1141鞭毛蛋白的特异性PTI诱导的分子机制，需要来自N1141或K1菌株的高度纯化的鞭毛蛋白。 在这里，我们描述了一种高质量的纯化方法。 avenae鞭毛蛋白，并用于PTI诱导研究。
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