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- To perform the swimming-induced paralysis, or SWIP assay, transfer L4 staged C. elegans larvae to a glass spot plate filled with liquid. Identify L4 hermaphrodites by the presence of the developing vulva. A half-moon-shaped patch at the midpoint of the body, and a whip-like tail.
To assess dopamine-mediated swimming, record the worm's movement characterized by C- shaped body bending and thrashing behavior. Usually, dopaminergic neurons release dopamine into the synaptic cleft, which in turn, binds dopamine receptors on the receiving cell.
Once the message is passed on, dopamine is released from the receptors, and unbound neurotransmitters can be taken up by the dopaminergic neuron through specialized transporter proteins and reused.
In larvae with altered dopamine signaling, caused by inactivating mutations in the dopamine transporter or drug treatments that increase dopamine release, dopamine accumulates in the synaptic cleft. Excess dopamine over stimulates the dopamine receptors interfering with the motor program and leaving the larvae paralyzed. Larvae that are unable to move will sink to the bottom of the plate.
In the example protocol, we will see detailed demonstrations of
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