Improve Research Reproducibility A Bio-protocol resource
Peer-reviewed
tooltip

Measuring Oxygen Consumption Rate in Caenorhabditis elegans

秀丽隐杆线虫中耗氧速率测定

Konstantinos Palikaras Konstantinos Palikaras
Nektarios Tavernarakis Nektarios Tavernarakis

发布: 2016年12月05日第6卷第23期 DOI: 10.21769/BioProtoc.2049 浏览次数: 11415

评审: Jyotiska ChaudhuriJian ChenAnonymous reviewer(s)

download pdf PDF PDF
ask Q&A
引用
收藏
Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of Nature, featuring study using the protocol.

May 2015

Advertisement

Special Issue on Protocols for Organoid Research
Technologies for RNA Detection
Presubmission Inquiry

实验方案合集

专注于特定技术或应用的、详细的、经过同行评审的实验方案合集

Cancer Research
Immunology
Developmental Biology
Microbiology
Neuroscience
Cell Imaging - A Special Collection for Cell Bio 2023
查看更多 more

相关实验方案

使用2'，7'-二氯二氢荧光素二乙酸盐测定秀丽隐杆线虫细胞内ROS

使用2'，7'-二氯二氢荧光素二乙酸盐测定秀丽隐杆线虫细胞内ROS

Dong Suk Yoon [...] Dong Seok Cha

2018年03月20日 17068 阅读

利用海马细胞能量代谢实时测定仪XF24 进行秀丽隐杆线虫中耗氧量测定

利用海马细胞能量代谢实时测定仪XF24 进行秀丽隐杆线虫中耗氧量测定

Suraiya Haroon and Marc Vermulst

2019年07月05日 6945 阅读

秀丽隐杆线虫荧光多聚体分析

秀丽隐杆线虫荧光多聚体分析

Dan Shaffer and Jarod A Rollins

2020年09月05日 7849 阅读

Abstract

The rate of oxygen consumption is a vital marker indicating cellular function during lifetime under normal or metabolically challenged conditions. It is used broadly to study mitochondrial function (Artal-Sanz and Tavernarakis, 2009; Palikaras et al., 2015; Ryu et al., 2016) or investigate factors mediating the switch from oxidative phosphorylation to aerobic glycolysis (Chen et al., 2015; Vander Heiden et al., 2009). In this protocol, we describe a method for the determination of oxygen consumption rates in the nematode Caenorhabditis elegans.

Keywords: Ageing (衰老) search Caenorhabditis elegans (秀丽隐杆线虫) search Metabolism (代谢) search Mitochondria (线粒体) search Oxygen sensor (氧传感器) search ROS (ROS) search

Background

Recent evidence underlines mitochondrial function as a potential contributor in the maintenance of organismal homeostasis and viability (Vafai and Mootha, 2012). Cellular oxygen consumption is highly recognized as a fundamental indicator of mitochondrial function, reflecting reactive oxygen species (ROS) production and metabolic activity of the cell. Therefore, several methods have been developed to measure oxygen consumption rates in cells or entire organisms (Dranka et al., 2011; Li and Graham, 2012; Luz et al., 2015; Perry et al., 2013). These approaches provided insight into the pivotal roles of mitochondria in disease progression and pathogenesis (Scheibye-Knudsen et al., 2015). In this protocol, we describe a method for the determination of oxygen consumption rates in the nematode C. elegans by using a Clark-type polarographic oxygen sensor electrode (Hansatech, King’s Lynn, England).

Materials and Reagents

  1. Consumables
    1. Commercial cigarette paper
    2. Polytetrafluorethylene (PTFE) membrane (provided by Hansatech, King’s Lynn, England)
    3. 15 ml tube (SARSTEDT, catalog number: 62.554.016 )
    4. 1.5 ml tube (Sigma-Aldrich, catalog number: Z606340 )
    5. Paper towel
    6. Greiner Petri dishes (60 x 15 mm) (Greiner Bio One, catalog number: 628161 )
    7. Maintenance kit (Hansatech, King’s Lynn, England)

  2. Biologicalreagents

    1. C. elegans strains (wild type [N2] and dct-1[tm376])
    2. Escherichia coli OP50 strain (obtained from the Caenorhabditis Genetics Center)

  3. Chemical reagents
    1. Distilled water
    2. Nitrogen gas provided in a tank
    3. PierceTM BCA Protein Assay Kit (Thermo Fisher Scientific, Thermo ScientificTM, catalog number: 23225 )
    4. Potassium dihydrogen phosphate (KH2PO4) (EMD Millipore, catalog number: 1048731000 )
    5. K2HPO4
    6. Sodium chloride (NaCl) (EMD Millipore, catalog number: 1064041000 )
    7. BactoTM peptone (BD, BactoTM, catalog number: 211677 )
    8. Streptomycin (Sigma-Aldrich, catalog number: S-6501 )
    9. Agar (Sigma-Aldrich, catalog number: 05040 )
    10. Cholesterol stock solution (SERVA Electrophoresis, catalog number: 17101.01 )
    11. Calcium chloride (CaCl2) (Sigma-Aldrich, catalog number: C-5080 )
    12. Magnesium sulfate (MgSO4) (Sigma-Aldrich, catalog number: M-7506 )
    13. Nystatin stock solution (Sigma-Aldrich, catalog number: N-3503 )
    14. Na2HPO4 (EMD Millipore, catalog number: 1065860500 )
    15. Potassium chloride (KCl) (Sigma-Aldrich, catalog number: P-5405 )
    16. Potassium chloride (KCl) buffer (see Recipes)
    17. Nematode growth medium (NGM) agar plates (see Recipes)
    18. M9 buffer (sterile; see Recipes)
    19. Phosphate buffer (sterile; see Recipes)

Equipment

  1. Dissecting stereomicroscope (Olympus, model: SMZ645 )
  2. Incubators for stable temperature (AQUA®LYTIC incubator 20 °C)
  3. DW1/AD Clark-type polarographic oxygen sensor (Hansatech Instruments, model: Oxygraph Plus System )
  4. Water bath at 20 °C
  5. Tabletop centrifuge (Eppendorf, model: 5424 )
  6. Sonicator (Sonics & Material, model: VC 130PB )

Software

  1. Oxygraph Plus software (Hansatech, King’s Lynn, England)
  2. Microsoft Office 2011 Excel

Procedure

English
中文翻译

文章信息

版权信息

© 2016 The Authors; exclusive licensee Bio-protocol LLC.

如何引用

Palikaras, K. and Tavernarakis, N. (2016). Measuring Oxygen Consumption Rate in Caenorhabditis elegans. Bio-protocol 6(23): e2049. DOI: 10.21769/BioProtoc.2049.

ris ris Download Citation in RIS Format

分类

发育生物学 > 细胞生长和命运决定 > 成熟衰老

生物化学 > 其它化合物 > 氧

您对这篇实验方法有问题吗？

在此处发布您的问题，我们将邀请本文作者来回答。同时，我们会将您的问题发布到Bio-protocol Exchange，以便寻求社区成员的帮助。

post 发布问题
0 Q&A
pen 提交稿件