搜索

The Method of the Body Bending Assay Using Caenorhabditis elegans
使用秀丽隐杆线虫进行身体弯曲试验法   

下载 PDF 引用 收藏 提问与回复 分享您的反馈 Cited by

本文章节

Abstract

This protocol is useful to obtain clear and repeatable data to know the motor function of a worm by counting the number of body thrash in M9 buffer. The thrashing assay is useful for observation of the effect of loss of motor neurons such as VA or VB neuron.

Materials and Reagents

  1. KH2PO4 (Wako Chemicals USA, catalog number:  169-04245 )
  2. Na2HPO4 (Wako Chemicals USA, catalog number:  197-02865 )
  3. NaCl (Wako Chemicals USA, catalog number:  191-01665 )
  4. M9 buffer (see Recipes)

Equipment

  1. Sterile NGM agar plate with a 35 mm diameter

Procedure

Fill sterile NGM agar plates with a 35 mm diameter with 1 ml of M9 buffer. Before the start of this assay, put a worm on a sterile NGM agar plate without bacteria and allow it to crawl freely to remove the agglomerated bacteria from the worm. After examining whether the bacteria were removed by visually, put a worm (4-day-old) into the buffer and allow it to swim freely for 1 min to be accustomed to the environment. 
Then count the number of thrash for 1 min (Nawa et al., 2012). Count at least 10 worms for each assay.
Note: A movement of the worm that swings its head and/or tail to the same side is counted as one thrash. A movie of body-bending worms makes counting more correct and easy. The bacteria that are served as food for worms should be removed as completely as possible because they affect the worm's movement.

Recipes

  1. M9 buffer (Brenner, 1974)

Acknowledgments

This protocol is adapted from Brenner (1974) and previously used in Nawa et al. (2012).

References

  1. Brenner, S. (1974). The genetics of Caenorhabditis elegans. Genetics 77(1): 71-94.
  2. Nawa, M., Kage-Nakadai, E., Aiso, S., Okamoto, K., Mitani, S. and Matsuoka, M. (2012). Reduced expression of BTBD10, an Akt activator, leads to motor neuron death. Cell Death Differ 19(8): 1398-1407.

简介

该协议通过计算M9缓冲器中主体thrash的数量来获得清楚和可重复的数据以了解蠕虫的运动功能是有用的。 颠簸测定法用于观察运动神经元如VA或VB神经元的损失的影响。

材料和试剂

  1. KH sub 2 PO 4(Wako Chemicals USA,目录号:169-04245)
  2. Na 2 HPO 4(Wako Chemicals USA,目录号:197-02865)
  3. NaCl(Wako Chemicals USA,目录号:191-01665)
  4. M9缓冲区(请参阅配方)

设备

  1. 具有35mm直径的无菌NGM琼脂板

程序

填充35毫米直径与1毫升M9缓冲液的无菌NGM琼脂板。在开始这种测定之前,将蠕虫放在无细菌的无菌NGM琼脂板上,并允许其自由爬行以从蠕虫中除去附聚的细菌。在检查细菌是否通过视觉去除后,将蠕虫(4天龄)放入缓冲液中,并允许其自由游动1分钟以习惯环境。
然后计数1分钟的瘙痒次数(Nawa等人,2012)。每个测定计数至少10个蠕虫。
注意:将其头部和/或尾部摆动到同一侧的蠕虫的运动被计为一个鞭毛。身体弯曲蠕虫的电影使计数更正确和容易。用作蠕虫食物的细菌应该尽可能完全地去除,因为它们会影响蠕虫的运动。

食谱

  1. M9缓冲区(Brenner,1974)

致谢

该协议改编自Brenner(1974)并且之前在Nawa等人(2012)中使用。

参考文献

  1. Brenner,S.(1974)。 Caenorhabditis elegans的遗传学 遗传学 77(1):71-94。
  2. Nawa,M.,Kage-Nakadai,E.,Aiso,S.,Okamoto,K.,Mitani,S。和Matsuoka,M。(2012)。 减少BTBD10(一种Akt激活剂)的表达,导致运动神经元死亡。 Cell Death Differ 19(8):1398-1407。
  • English
  • 中文翻译
免责声明 × 为了向广大用户提供经翻译的内容,www.bio-protocol.org 采用人工翻译与计算机翻译结合的技术翻译了本文章。基于计算机的翻译质量再高,也不及 100% 的人工翻译的质量。为此,我们始终建议用户参考原始英文版本。 Bio-protocol., LLC对翻译版本的准确性不承担任何责任。
Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.
引用:Nawa, M. and Matsuoka, M. (2012). The Method of the Body Bending Assay Using Caenorhabditis elegans. Bio-protocol 2(17): e253. DOI: 10.21769/BioProtoc.253.
提问与回复

(提问前,请先登录)bio-protocol作为媒介平台,会将您的问题转发给作者,并将作者的回复发送至您的邮箱(在bio-protocol注册时所用的邮箱)。为了作者与用户间沟通流畅(作者能准确理解您所遇到的问题并给与正确的建议),我们鼓励用户用图片或者视频的形式来说明遇到的问题。由于本平台用Youtube储存、播放视频,作者需要google 账户来上传视频。

当遇到任务问题时,强烈推荐您提交相关数据(如截屏或视频)。由于Bio-protocol使用Youtube存储、播放视频,如需上传视频,您可能需要一个谷歌账号。