An imaging system for standardized quantitative analysis of C. elegans behavior

Feng, Zhaoyang; Cronin, Christopher J.; Wittig, John H.; Sternberg, Paul W.; Schafer, William R.

doi:10.1186/1471-2105-5-115

Cited by 108 publications

(73 citation statements)

References 11 publications

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“…Since the original description of the swimming (thrashing) assay by Epstein and Thomson in 1974 [16], a number of other methods mostly involving video recording and computer/mathematical analysis for quantifying C. elegans motility have appeared in the literature [43–49]. However, most of these methods are designed to analyze complex behaviors such as foraging, chemotaxis and response to vibration, and were not specifically designed or applied to assess muscle mutants.…”

Section: Resultsmentioning

confidence: 99%

Bending amplitude – A new quantitative assay of C. elegans locomotion: Identification of phenotypes for mutants in genes encoding muscle focal adhesion components

Nahabedian

Qadota

Stirman

et al. 2012

Methods

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The nematode Caenorhabditis elegans uses striated muscle in its body wall for locomotion. The myofilament lattice is organized such that all the thin filament attachment structures (dense bodies, analogous to Z-disks) and thick filament organizing centers (M-lines) are attached to the muscle cell membrane. Thus, the force of muscle contraction is transmitted through these structures and allows locomotion of the worm. Dense bodies and M-lines are compositionally similar to focal adhesions and costameres, and are based on integrin and associated proteins. Null mutants for many of the newly discovered dense body and M-line proteins do not have obvious locomotion defects when observed casually, or when assayed by counting the number of times a worm moves back and forth in liquid. We hypothesized that many of these proteins, located as they are in muscle focal adhesions, function in force transmission, but we had not used an appropriate or sufficiently sensitive assay to reveal this function. Recently, we have developed a new quantitative assay of C. elegans locomotion that measures the maximum bending amplitude of an adult worm as it moves backwards. The assay had been used to reveal locomotion defects for null mutants of genes encoding ATN-1 (α-actinin) and PKN-1 (protein kinase N). Here, we describe the details of this method, and apply it to 21 loss of function mutants in 17 additional genes, most of which encode components of muscle attachment structures. As compared to wild type, mutations in 11 genes were found to have less ability to bend, and mutations in one gene were found to have greater ability to bend. Loss of function mutants for eight proteins had been reported to have normal locomotion (ZYX-1(zyxin), ALP-1 (Enigma), DIM-1, SCPL-1), or locomotion that was not previously investigated (FRG-1 (FRG1), KIN-32 (focal adhesion kinase, LIM-8), or had only slightly decreased locomotion (PFN-3 (profilin)).

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Section: Resultsmentioning

confidence: 99%

Bending amplitude – A new quantitative assay of C. elegans locomotion: Identification of phenotypes for mutants in genes encoding muscle focal adhesion components

Nahabedian

Qadota

Stirman

et al. 2012

Methods

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“…When deciding what parameters were to be quantified or characterized by Track-A-Worm , we referred to parameters found in several leading worm trackers [1], [2], [5] as well information available about their current versions at the developers' websites (http://wormlab.caltech.edu/publications/download.html; http://www.mrc-lmb.cam.ac.uk/wormtracker/), and considered the potential common needs of C. elegans research labs. The current version of Track-A-Worm allows the quantification of most of the parameters found in the other systems.…”

Section: Discussionmentioning

confidence: 99%

“…However, there has not been a single-worm tracker with wide implementations. The Sternberg lab and the Schafer lab were among the first developing feature-rich single-worm trackers [1], [2]. The hardware designs and software, including subsequent updates, have been available to the public (http://wormlab.caltech.edu/publications/download.html; http://www.mrc-lmb.cam.ac.uk/wormtracker/).…”

Section: Introductionmentioning

confidence: 99%

Track-A-Worm, An Open-Source System for Quantitative Assessment of C. elegans Locomotory and Bending Behavior

Wang

2013

PLoS ONE

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A major challenge of neuroscience is to understand the circuit and gene bases of behavior. C. elegans is commonly used as a model system to investigate how various gene products function at specific tissue, cellular, and synaptic foci to produce complicated locomotory and bending behavior. The investigation generally requires quantitative behavioral analyses using an automated single-worm tracker, which constantly records and analyzes the position and body shape of a freely moving worm at a high magnification. Many single-worm trackers have been developed to meet lab-specific needs, but none has been widely implemented for various reasons, such as hardware difficult to assemble, and software lacking sufficient functionality, having closed source code, or using a programming language that is not broadly accessible. The lack of a versatile system convenient for wide implementation makes data comparisons difficult and compels other labs to develop new worm trackers. Here we describe Track-A-Worm, a system rich in functionality, open in source code, and easy to use. The system includes plug-and-play hardware (a stereomicroscope, a digital camera and a motorized stage), custom software written to run with Matlab in Windows 7, and a detailed user manual. Grayscale images are automatically converted to binary images followed by head identification and placement of 13 markers along a deduced spline. The software can extract and quantify a variety of parameters, including distance traveled, average speed, distance/time/speed of forward and backward locomotion, frequency and amplitude of dominant bends, overall bending activities measured as root mean square, and sum of all bends. It also plots worm travel path, bend trace, and bend frequency spectrum. All functionality is performed through graphical user interfaces and data is exported to clearly-annotated and documented Excel files. These features make Track-A-Worm a good candidate for implementation in other labs.

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“…For this work we show that worms transition between forward and backward states preferentially at different points in its undulatory cycle, a measure that is difficult to do by eye. This work is complementary to tracking systems that have been developed to measure behavior of worms at lower magnification 8,9,10 , and also to systems that are able to measure or modulate neuronal activity using genetically encoded reagents 11,12 . While single organism tracking is a powerful method for quantification of behavior there are some limitations to the technique.…”

Section: Discussionmentioning

confidence: 99%

<em>C. elegans</em> Tracking and Behavioral Measurement

Likitlersuang

Stephens

Palanski

et al. 2012

JoVE

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We have developed instrumentation, image processing, and data analysis techniques to quantify the locomotory behavior of C. elegans as it crawls on the surface of an agar plate. For the study of the genetic, biochemical, and neuronal basis of behavior, C. elegans is an ideal organism because it is genetically tractable, amenable to microscopy, and shows a number of complex behaviors, including taxis, learning, and social interaction. Behavioral analysis based on tracking the movements of worms as they crawl on agar plates have been particularly useful in the study of sensory behavior, locomotion, and general mutational phenotyping. Our system works by moving the camera and illumination system as the worms crawls on a stationary agar plate, which ensures no mechanical stimulus is transmitted to the worm. Our tracking system is easy to use and includes a semi-automatic calibration feature. A challenge of all video tracking systems is that it generates an enormous amount of data that is intrinsically high dimensional. Our image processing and data analysis programs deal with this challenge by reducing the worms shape into a set of independent components, which comprehensively reconstruct the worms behavior as a function of only 3-4 dimensions. As an example of the process we show that the worm enters and exits its reversal state in a phase specific manner.

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An imaging system for standardized quantitative analysis of C. elegans behavior

Cited by 108 publications

References 11 publications

Bending amplitude – A new quantitative assay of C. elegans locomotion: Identification of phenotypes for mutants in genes encoding muscle focal adhesion components

Bending amplitude – A new quantitative assay of C. elegans locomotion: Identification of phenotypes for mutants in genes encoding muscle focal adhesion components

Track-A-Worm, An Open-Source System for Quantitative Assessment of C. elegans Locomotory and Bending Behavior

<em>C. elegans</em> Tracking and Behavioral Measurement

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