A Method to Injure, Dissect and Image Indirect Flight Muscle of Drosophila
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Author:
Date:
2018-05-20
[Abstract] Inducing an injury specifically to Drosophila flight muscles is a difficult task, owing to the small size of the muscles and the presence of the cuticle. The protocol described below provides an easy and reproducible method to induce injury in the Drosophila flight muscles.
[摘要] 由于肌肉尺寸小和角质层的存在,特别针对果蝇飞行肌肉诱导损伤是一项艰巨的任务。 下面描述的方案提供了一种简单且可重复的方法来诱导果蝇飞行肌肉中的损伤。
【背景】脊椎动物的肌肉进行再生,这是一种归因于卫星细胞(即常驻干细胞)的过程。 我们的实验室最近表明,果蝇的肌肉具有与脊椎动物卫星细胞相似的干细胞,即昆虫卫星细胞,并显示对肌肉损伤的增殖反应(Chaturvedi等人,2017年))。 飞蝇遗传学的易用性和我们的诱导伤害的方法为解决再生生物学领域的相关问题提供了机会。 我们已经标准化了一种以更好的精确度损伤背侧纵肌(DLMs)纤维的方案,并且该方法可以用于研究损伤后肌肉中涉及的修复机制。
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In Gel Kinase Assay
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Author:
Date:
2017-03-05
[Abstract] Proper spatiotemporal regulation of protein phosphorylation in cells and tissues is required for normal development and homeostasis. We present the protocol ‘In Gel Kinase Assay’, which is useful for protein kinase activity measurements from crude protein extracts. We have successfully used ‘In Gel Kinase Assay’ protocol to show that the Arabidopsis thaliana sextuple mutant in the PYRABACTIN RESISTANCE1/PYR1-LIKE/REGULATORY COMPONENTS OF ABA RECEPTORS (PYR/PYL/RCAR-ABA receptors; line pyr/pyl112458) is impaired in ABA-mediated activation of SnRK2.2, SnRK2.3 and OST1/SnRK2.6, ...
[摘要] 正常发育和体内平衡需要细胞和组织中蛋白质磷酸化的适时时空调节。我们提出方案“凝胶激酶测定”,其可用于粗蛋白质提取物的蛋白激酶活性测量。我们已经成功地使用“凝胶激酶测定”方案来证明在ABA受体(PYR / PYL / RCAR-ABA受体)的PYRABACTIN RESISTANCE1 / PYR1-样/调节组分中的拟南芥线条pyr / pyl112458 )在ABA介导的SnRK2.2,SnRK2.3和OST1 / SnRK2.6的活化中受损,多达三重突变体snrk2.2 / 2.3 / 2.6 (Gonzalez-Guzman等人,2012)。
背景 植物激素脱落酸(ABA)是涉及植物生长发育以及植物对非生物和生物胁迫的反应的关键信号。 ABA感知和信号通路由ABA受体(PYR / PYL / RCAR-ABA受体)的PYRABACTIN RESISTANCE1 / PYR1-调节组分,PP2C磷酸酶和SnRK2s激酶组成(在Antoni等人, ,,2011)。模块受体-ABA-磷酸酶通过调节ABA激活的SnRK2而以配体依赖的方式控制磷酸化信号级联。反过来,SnRK2s激酶使细胞核和细胞质中的无数效应物从转录因子(例如,ABFs)到离子通道(例如)磷酸化, ...
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Atomic Force Microscopy (AFM) Analysis of Cell Wall Structural Glycoproteins in vitro
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Author:
Date:
2015-07-20
[Abstract] Hydroxyproline-rich glycoproteins (HRGPs) are major protein components in dicot primary cell walls and generally account for more than 10% of the wall dry weight. As essential members of the HRGP superfamily, extensins (EXTs) presumably function in the cell wall by assembling into positively charged protein scaffolds (Cannon et al., 2008) that direct the proper deposition of other wall polysaccharides, especially pectins, to ensure correct cell wall assembly (Hall and Cannon, 2002; Lamport et al., 2011a). Extensins are recalcitrant to purification as they are rapidly ...
[摘要] 富含羟脯氨酸的糖蛋白(HRGP)是双子叶原代细胞壁中的主要蛋白质组分,通常占壁干重的10%以上。作为HRGP超家族的主要成员,通过装配到带正电荷的蛋白质支架(Cannon等人,2008)中,可以推测伸展蛋白(EXT)在细胞壁中起作用,其指导其他壁多糖的适当沉积,特别是果胶,以确保正确的细胞壁组装(Hall和Cannon,2002; Lamport等人,2011a)。延伸素难以纯化,因为它们在进入细胞壁后快速交联成共价网络,但是存在可以提取新合成的延伸蛋白单体的短时间窗(Smith等人, 1984; Smith等人,,1986)。用于提取延伸蛋白和其他壁结构蛋白的详细方案早已被描述(Lamport等人,2011b)。这里详述的方案提供了一种方法,用于使用AFM在体外研究延伸素和潜在地其他细胞壁组分的自组装。
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