This video introduces the touch habituation assay, a C. elegans behavioral method of assessing mechanosensation and quantifying time to habituation.
The following protocol is an excerpt from Mersha et al. Long-term Behavioral and Reproductive Consequences of Embryonic Exposure to Low-dose Toxicants, J. Vis. Exp. (2018)
- Anterior Touch Habituation Assay
- Transfer approximately 10 synchronized young adult worms to new unseeded NGM plates using a fire-sterilized 30 G platinum wire pick. Leave worms undisturbed for 5 min to allow them time to acclimatize to the new plate.
- For the habituation response, use an eyebrow hair attached to the end of a wooden skewer or toothpick, and sterilize by dipping in 70% ethanol. Wipe with a clean lint-free tissue and wait for 1 min for ethanol to evaporate off. Gently touch the worm on the head (anterior of the pharyngeal bulb) using the eyebrow hair. Repeat the touches, allowing 10 s in between touches in order to allow the worm to recover.
- Continue to touch (allowing 10 s interstimulus intervals) until the worm no longer moves backwards. At this point, the worm has habituated to the stimulus. Record the num+ber of touches required for the worm to habituate.
- Analyze the difference between the mean habituation rates of the treated and untreated controls using one-way ANOVA followed by Tukey's post hoc test.
Subscription Required. Please recommend JoVE to your librarian.
|N2 (Wild-type C. elegans) worms||CGC||Repository at U of Minnesota|
|Platinum wire pick||Genesee Scientific||59-AWP|