Total Triglyceride Quantification in Caenorhabditis elegans
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Author:
Date:
2020-11-20
[Abstract] Several studies suggest an important role of lipid metabolism in regulating longevity of Caenorhabditis elegans. Therefore, assays to quantify lipids have enormous value in understanding aging and pathologies associated with it. Approximately 70% of lipid metabolism genes in the nematode have orthologs in humans. Amenability of C. elegans to genetic manipulations has allowed investigations into the role of specific genetic factors in lipid metabolism. Here, we describe a protocol to quantify total triglycerides in C. elegans, which can be extended to studies of the effects of altered environmental and genetic factors on stored fats. This protocol quantifies the picomoles of the triglycerides, in whole worm lysate. Due to the sensitivity of the assay, it could ...
[摘要] [摘要]多项研究表明脂质代谢在调节秀丽隐杆线虫的寿命中具有重要作用。因此,定量脂质的测定在理解衰老和与其相关的病理学方面具有巨大价值。线虫中大约70%的脂质代谢基因在人类中具有直系同源物。秀丽隐杆线虫对基因操作的适应性允许调查特定遗传因素在脂质代谢中的作用。在这里,我们描述了一个协议,以定量总甘油三酯在秀丽隐杆线虫,这CA n可以扩展到环境和遗传因素改变对储存脂肪的影响的研究。该协议量化了整个蠕虫裂解物中甘油三酸酯的皮摩尔。由于测定的敏感性,它可以帮助鉴定总存储脂肪的细微变化,这是显微镜技术无法识别的。
[背景]多余的卡路里通常以三酰基甘油或糖原的形式存储(Cohen,2018年)。甘油三酸酯(TAGs)的异常水平与人类的心脏病,胰腺炎和动脉粥样硬化有关(Cameron等人,1974; Lee等人,2003; Walther和Farese,2012; Onal等人,2017)。TAG被包装到细胞质中的脂滴中。脂质液滴存在于真核细胞中,大小从20 nm到100 µm不等(Stobart等,1986; ...
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A Quick Method for Screening Biocontrol Efficacy of Bacterial Isolates against Bacterial Wilt Pathogen Ralstonia solanacearum in Tomato
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Author:
Date:
2020-11-20
[Abstract] Ralstonia solanacearum is a bacterial phytopathogen able to cause bacterial wilt disease in more than 200 plant species. Plant disease biocontrol strategies are used for controlling this disease and tomato is used as a model plant to conduct R. solanacearum associated studies. Conventional screening methods such as seed bacterization, soil drenching and root bacterization (in grown plants) to assess the ability of biocontrol bacteria to antagonize R. solanacearum under in planta conditions in different hosts are time-consuming and costly. A fast, cost effective method is a key requirement to advance the research on R. solanacearum biocontrol. In this protocol, we have inoculated the roots of tomato seedlings with bacterial isolates showing antagonistic activity against R. solanacearum ...
[摘要] [摘要]青枯雷尔氏菌是能够导致青枯病在超过2个细菌植物病原体00植物物种。植物病害生物防治策略用于控制这种疾病和番茄被用来作为模式植物进行青枯菌相关的研究。常规的筛选方法,例如种子杀菌,土壤浸湿和根部杀菌(在生长中的植物中),以评估生物控制细菌在不同宿主中在植物条件下拮抗青枯菌的能力,既耗时又昂贵。一个快速,经济有效的方法是推动研究的关键要求 青枯菌生物防治。在此协议中,我们已在细菌的分离株上接种了番茄幼苗的根,这些分离株在体外条件下显示出对茄青枯菌的拮抗活性。用拮抗细菌处理16小时后,通过成熟的根浸法将青枯菌接种到幼苗中。然后将幼苗保持在受控条件和枯萎/死苗的数目,记录最多10个天交ř 。青枯菌接种。从每个测试的分离物的记录计算生物防治效力。此协议是比在感测已经可用的协议的优点在于,它可以在很短的持续时间内完成(〜18天番茄)并且没有保持苗培养介质的需求。该方法可用于在短时间内以最小的成本快速筛选大量细菌分离株和不同宿主基因型。
[背景[ Ralstonia solanacearum ]是一种土壤传播的细菌性植物病原体,在200多个农作物中引起细菌性枯萎病,代表50个不同的科(Seleim等人,2014),其中大部分来自茄科(Thera等人,2010)。它可以在土壤中壮成长长达数年之久,并具有在寄主到达时感染的能力(Coutinho and ...
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Pentylenetetrazole (PTZ)-induced Convulsion Assay to Determine GABAergic Defects in Caenorhabditis elegans
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Author:
Date:
2018-09-05
[Abstract] Pentylenetetrazole (PTZ) is a GABAA receptor antagonist and is used to monitor presynaptic defects in the release of the inhibitory neurotransmitter GABA. PTZ is a competitive inhibitor of GABA, and prevents binding of GABA on the GABAA receptors present on the surface of muscle. In the absence of GABA binding, the excitatory to inhibitory signal ratio increases resulting in a convulsive phenotype. This assay provides a fast and reliable method to detect presynaptic defects in GABAergic synaptic transmission. The assay is based on correlating the extent of convulsions with the degree of presynaptic GABA release defects.
[摘要] 戊四唑(PTZ)是GABA A 受体拮抗剂,用于监测抑制性神经递质GABA释放中的突触前缺陷。 PTZ是GABA的竞争性抑制剂,并且阻止GABA与肌肉表面上存在的GABA A 受体结合。 在没有GABA结合的情况下,抑制信号比的兴奋性增加导致惊厥表型。 该试验提供了一种快速可靠的方法来检测GABA能突触传递中的突触前缺陷。 该测定基于将惊厥的程度与突触前GABA释放缺陷的程度相关联。
【背景】突触是介导突触传递的不对称细胞间连接。它们构成了能够执行复杂行为的大脑电路的基本单元。动作电位的到达通过电压门控钙通道引起钙流入,导致质膜的去极化。在这种突触前去极化后,突触囊泡与质膜融合,通过高度控制的过程将其内容物释放到突触间隙中(Sudhof,1995; Sudhof,2004)。
运动是 Caenorhabditis elegans 中突出的行为输出。产生协调的背腹窦弯曲的神经回路允许 C中的正常运动。线虫。运动行为在多个层面上进行协调,并且涉及由中间神经元处理的多个感觉神经元整合多种感觉线索,并最终在神经肌肉接头(NMJ)处直接变化(de Bono和Maricq,2005; Bargmann,2012)。 C.线虫 NMJ有三个主要组成部分;兴奋性胆碱能运动神经元,抑制性GABA能神经运动神经元和突触后肌细胞(White et ...
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