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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.

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The Method of the Body Bending Assay Using Caenorhabditis elegans

Neuroscience > Behavioral neuroscience > Cognition
Authors: Mikiro Nawa
Mikiro Nawa Affiliation: Department of Pharmacology, Tokyo Medical University, Shinjuku-ku, Tokyo, Japan
Bio-protocol author page: a89
 and Masaaki Matsuoka
Masaaki MatsuokaAffiliation: Department of Pharmacology, Tokyo Medical University, Shinjuku-ku, Tokyo, Japan
For correspondence: sakimatu@tokyo-med.ac.jp
Bio-protocol author page: a90
Vol 2, Iss 17, 9/5/2012, 5909 views, 0 Q&A, How to cite
DOI: http://dx.doi.org/10.21769/BioProtoc.253

[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.


How to cite this protocol: 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; Full Text



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