C. elegans Chemotaxis Tracking
Objective: To investigate the behavioral mechanism of chemotaxis in C. elegans by recording instantaneous position, speed, and turning rate during chemotaxis in chemical gradients
Gather these items before starting the experiment. Check off items as you prepare.
Equipment1
Not specified • Not specified • Not specified • Not mentioned
Materials2
Software1
Not specified • Not mentioned
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Protocol Steps
Prepare chemical gradient
Create gradients of attractant chemicals (ammonium chloride or biotin) for chemotaxis assay
Note: Two different attractants were tested
View evidence from paper
“chemotaxis in gradients of the attractants ammonium chloride or biotin”
Place single worm in gradient
Position a single C. elegans worm in the chemical gradient environment
Note: Single worms were tracked individually
View evidence from paper
“recorded the instantaneous position, speed, and turning rate of single worms as a function of time during chemotaxis”
Record worm movement
Record instantaneous position, speed, and turning rate of the worm as a function of time during chemotaxis
Note: Continuous tracking of movement parameters
View evidence from paper
“recorded the instantaneous position, speed, and turning rate of single worms as a function of time during chemotaxis”
Analyze turning rate patterns
Divide worm tracks into periods of smooth swimming (runs) and periods of frequent turning (pirouettes) based on turning rate analysis
Note: Two distinct behavioral states identified
View evidence from paper
“Analysis of turning rate showed that each worm track could be divided into periods of smooth swimming (runs) and periods of frequent turning (pirouettes)”
Correlate pirouettes with concentration gradient
Analyze the correlation between pirouette initiation and the rate of change of concentration (dC/dt) rather than absolute concentration
Note: Pirouettes correlated with dC/dt, not absolute concentration
View evidence from paper
“The initiation of pirouettes was correlated with the rate of change of concentration (d C /d t ) but not with absolute concentration”
Determine pirouette directional bias
Analyze the probability of pirouette occurrence relative to worm heading direction in the gradient
Note: Pirouettes most likely when heading down gradient, least likely when heading up gradient
View evidence from paper
“Pirouettes were most likely to occur when a worm was heading down the gradient (d C /d t < 0) and least likely to occur when a worm was heading up the gradient (d C /d t > 0)”
Analyze post-pirouette direction
Determine the average direction of movement after a pirouette occurs
Note: Average direction of movement after pirouette was up the gradient
View evidence from paper
“Further analysis revealed that the average direction of movement after a pirouette was up the gradient”
Test pirouette model with stochastic simulation
Impose the correlation between pirouettes and dC/dt on a stochastic point model of worm motion to test if the model exhibits chemotaxis behavior
Note: Model tested in both radial and planar gradients
View evidence from paper
“We tested this idea by imposing the correlation between pirouettes and d C /d t on a stochastic point model of worm motion. The model exhibited chemotaxis behavior in a radial gradient and also in a novel planar gradient”