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Locomotory Rate Assay, C. elegans
Author: Sanjib Guha, updated date: , view: 27, Q&A: 0
Tags: Worm and Locomotion

Materials and Reagents

Small plates.

Age synchronize animals.

Fresh OP50.

Kimwipes.

Timer.

Video equipment, if needed.


Procedure

Assay plates were prepared by spreading the E. coli strain OP50-1 in a ring on NGM agar (Brenner 1974) in 5 cm petri plates.

Assay plates were always freshly spread with bacteria, incubated overnight at 37°C, and allowed to cool to room temperature before use.

Plates for measuring locomotory rate in the absence of bacteria were also incubated at 37°C. Only synchronized young adult hermaphrodites (16 h after the late L4 larval stage) were tested.

In all cases, plates were coded so that the experimenter was blind to the genotype.

For well-fed animals, locomotory rate was measured by removing 5 animals from plates with ample bacteria, washing the animals twice in S basal buffer (Brenner 1974), and transferring them to an assay plate in a drop of buffer using a capillary pipette.

The drop of buffer used to transfer the animals was absorbed with a Kimwipe.

Five minutes after transfer, the number of body bends in 20 s intervals was sequentially counted for each of the 5 animals on the assay plate and then repeated the same thing for next set of animals in a different assay plate.

References

Sawin E. R., Ranganathan R. & Horvitz H. R. C. elegans locomotory rate is modulated by the environment through a dopaminergic pathway and by experience through a serotonergic pathway. Cell press 26: 619-631 (2000).


Brenner S. The genetics of Caenorhabtiditis elegans. Genetics 77: 71-94 (1974).


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