The Parallel Worm Tracker: A Platform for Measuring Average Speed and Drug-Induced Paralysis in Nematodes

Background Caenorhabditis elegans locomotion is a simple behavior that has been widely used to dissect genetic components of behavior, synaptic transmission, and muscle function. Many of the paradigms that have been created to study C. elegans locomotion rely on qualitative experimenter observation. Here we report the implementation of an automated tracking system developed to quantify the locomotion of multiple individual worms in parallel. Methodology/Principal Findings Our tracking system generates a consistent measurement of locomotion that allows direct comparison of results across experiments and experimenters and provides a standard method to share data between laboratories. The tracker utilizes a video camera attached to a zoom lens and a software package implemented in MATLAB®. We demonstrate several proof-of-principle applications for the tracker including measuring speed in the absence and presence of food and in the presence of serotonin. We further use the tracker to automatically quantify the time course of paralysis of worms exposed to aldicarb and levamisole and show that tracker performance compares favorably to data generated using a hand-scored metric. Conclusions/Signficance Although this is not the first automated tracking system developed to measure C. elegans locomotion, our tracking software package is freely available and provides a simple interface that includes tools for rapid data collection and analysis. By contrast with other tools, it is not dependent on a specific set of hardware. We propose that the tracker may be used for a broad range of additional worm locomotion applications including genetic and chemical screening.

[1]  H. Spencer The structure of the nervous system. , 1870 .

[2]  Hilla Peretz,et al.  Ju n 20 03 Schrödinger ’ s Cat : The rules of engagement , 2003 .

[3]  S. Brenner The genetics of Caenorhabditis elegans. , 1974, Genetics.

[4]  J. Lewis,et al.  The genetics of levamisole resistance in the nematode Caenorhabditis elegans. , 1980, Genetics.

[5]  James A. Lewis,et al.  THE GENETICS OF LEVAMISOLE RESISTANCE IN THE , 1980 .

[6]  D B Dusenbery,et al.  Using a microcomputer and video camera to simultaneously track 25 animals. , 1985, Computers in biology and medicine.

[7]  S. Brenner,et al.  The structure of the nervous system of the nematode Caenorhabditis elegans. , 1986, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.

[8]  J. Fleming,et al.  Cholinergic receptor mutants of the nematode Caenorhabditis elegans , 1987, Journal of Neuroscience.

[9]  Edward H. Coe,et al.  The Genetics of Corn , 1988 .

[10]  R. Hosono,et al.  Additional genes which result in an elevation of acetylcholine levels by mutations in Caenorhabditis elegans , 1991, Neuroscience Letters.

[11]  M. Nonet,et al.  Synaptic function is impaired but not eliminated in C. elegans mutants lacking synaptotagmin , 1993, Cell.

[12]  C. Johnson,et al.  Caenorhabditis elegans mutants resistant to inhibitors of acetylcholinesterase. , 1995, Genetics.

[13]  J A Crowell,et al.  A genetic selection for Caenorhabditis elegans synaptic transmission mutants. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[14]  William R. Schafer,et al.  Control of Alternative Behavioral States by Serotonin in Caenorhabditis elegans , 1998, Neuron.

[15]  S. Lockery,et al.  Active Currents Regulate Sensitivity and Dynamic Range in C. elegans Neurons , 1998, Neuron.

[16]  Cori Bargmann,et al.  Natural Variation in a Neuropeptide Y Receptor Homolog Modifies Social Behavior and Food Response in C. elegans , 1998, Cell.

[17]  M. Futai,et al.  Sensing of cadmium and copper ions by externally exposed ADL, ASE, and ASH neurons elicits avoidance response in Caenorhabditis elegans. , 1999, Neuroreport.

[18]  A. V. Maricq,et al.  Neuronal Control of Locomotion in C. elegans Is Modified by a Dominant Mutation in the GLR-1 Ionotropic Glutamate Receptor , 1999, Neuron.

[19]  Thomas M. Morse,et al.  The Fundamental Role of Pirouettes in Caenorhabditis elegans Chemotaxis , 1999, The Journal of Neuroscience.

[20]  J. Thomas,et al.  Calcium/calmodulin-dependent protein kinase II regulates Caenorhabditis elegans locomotion in concert with a G(o)/G(q) signaling network. , 2000, Genetics.

[21]  Ryuzo Shingai,et al.  Durations and frequencies of free locomotion in wild type and GABAergic mutants of Caenorhabditis elegans , 2000, Neuroscience Research.

[22]  Rajesh Ranganathan,et al.  C. elegans Locomotory Rate Is Modulated by the Environment through a Dopaminergic Pathway and by Experience through a Serotonergic Pathway , 2000, Neuron.

[23]  R. Kerr,et al.  Serotonin modulates locomotory behavior and coordinates egg-laying and movement in Caenorhabditis elegans. , 2001, Journal of neurobiology.

[24]  E. Cuppen,et al.  The G-protein beta-subunit GPB-2 in Caenorhabditis elegans regulates the G(o)alpha-G(q)alpha signaling network through interactions with the regulator of G-protein signaling proteins EGL-10 and EAT-16. , 2001, Genetics.

[25]  Lawrence Salkoff,et al.  SLO-1 Potassium Channels Control Quantal Content of Neurotransmitter Release at the C. elegans Neuromuscular Junction , 2001, Neuron.

[26]  Motomichi Doi,et al.  Regulation of Retrograde Signaling at Neuromuscular Junctions by the Novel C2 Domain Protein AEX-1 , 2002, Neuron.

[27]  P. Sengupta,et al.  Regulation of Body Size and Behavioral State of C. elegans by Sensory Perception and the EGL-4 cGMP-Dependent Protein Kinase , 2002, Neuron.

[28]  P. Cosman,et al.  Using machine vision to analyze and classify Caenorhabditis elegans behavioral phenotypes quantitatively , 2002, Journal of Neuroscience Methods.

[29]  Aravinthan D. T. Samuel,et al.  Thermotaxis in Caenorhabditis elegans Analyzed by Measuring Responses to Defined Thermal Stimuli , 2002, The Journal of Neuroscience.

[30]  W. Wadsworth,et al.  The C Domain of Netrin UNC-6 Silences Calcium/Calmodulin-Dependent Protein Kinase- and Diacylglycerol-Dependent Axon Branching in Caenorhabditis elegans , 2002, The Journal of Neuroscience.

[31]  J. Kaplan,et al.  The EGL-21 Carboxypeptidase E Facilitates Acetylcholine Release at Caenorhabditis elegans Neuromuscular Junctions , 2003, The Journal of Neuroscience.

[32]  D. Hall,et al.  Long chain polyunsaturated fatty acids are required for efficient neurotransmission in C. elegans , 2003, Journal of Cell Science.

[33]  Pamela C. Cosman,et al.  Quantitative Classification and Natural Clustering of C. elegans Behavioral Phenotypes , 2003 .

[34]  P. Cosman,et al.  Quantitative classification and natural clustering of Caenorhabditis elegans behavioral phenotypes. , 2003, Genetics.

[35]  M. Nonet,et al.  Resistance to Volatile Anesthetics by Mutations Enhancing Excitatory Neurotransmitter Release in Caenorhabditis elegans , 2004, Genetics.

[36]  Pamela C. Cosman,et al.  Automatic tracking, feature extraction and classification of C. elegans phenotypes , 2004, IEEE Transactions on Biomedical Engineering.

[37]  Paul W. Sternberg,et al.  An imaging system for standardized quantitative analysis of C. elegans behavior , 2004, BMC Bioinformatics.

[38]  Oliver Hobert,et al.  A Conserved Postsynaptic Transmembrane Protein Affecting Neuromuscular Signaling in Caenorhabditis elegans , 2004, The Journal of Neuroscience.

[39]  Christopher J. Cronin,et al.  An automated system for measuring parameters of nematode sinusoidal movement , 2005, BMC Genetics.

[40]  Phillip L. Williams,et al.  The nematode Caenorhabditis elegans as a model of organophosphate-induced mammalian neurotoxicity. , 2004, Toxicology and applied pharmacology.

[41]  J. Richmond Synaptic function. , 2005, WormBook : the online review of C. elegans biology.

[42]  Marc Vidal,et al.  Systematic analysis of genes required for synapse structure and function , 2005, Nature.

[43]  K. Miller,et al.  Convergent, RIC-8-Dependent Gα Signaling Pathways in the Caenorhabditis elegans Synaptic Signaling Network , 2005, Genetics.

[44]  Phillip L. Williams,et al.  Using Transgenic Caenorhabditis elegans in Soil Toxicity Testing , 2005, Archives of environmental contamination and toxicology.

[45]  Mario de Bono,et al.  Experience-Dependent Modulation of C. elegans Behavior by Ambient Oxygen , 2005, Current Biology.

[46]  S. Lockery,et al.  Analysis of the effects of turning bias on chemotaxis in C. elegans , 2005, Journal of Experimental Biology.

[47]  S. Nurrish,et al.  Rho is a presynaptic activator of neurotransmitter release at pre-existing synapses in C. elegans. , 2006, Genes & development.

[48]  John M. Walker,et al.  C. elegans , 2006, Methods in Molecular Biology.

[49]  Leon Avery,et al.  Dietary choice behavior in Caenorhabditis elegans , 2006, Journal of Experimental Biology.

[50]  Theresa Stiernagle Maintenance of C. elegans. , 2006, WormBook : the online review of C. elegans biology.

[51]  S. Nurrish,et al.  The Gα12‐RGS RhoGEF‐RhoA signalling pathway regulates neurotransmitter release in C. elegans , 2006, The EMBO journal.

[52]  Christopher J. Cronin,et al.  Conservation rules, their breakdown, and optimality in Caenorhabditis sinusoidal locomotion. , 2006, Journal of theoretical biology.

[53]  Nektarios Tavernarakis,et al.  Caenorhabditis elegans: A versatile platform for drug discovery , 2006, Biotechnology journal.

[54]  P. Cosman,et al.  Machine vision based detection of omega bends and reversals in C. elegans , 2006, Journal of Neuroscience Methods.

[55]  Timothy R Mahoney,et al.  Analysis of synaptic transmission in Caenorhabditis elegans using an aldicarb-sensitivity assay , 2006, Nature Protocols.

[56]  Joel Burdick,et al.  Automated Tracking of Multiple C. Elegans , 2006, 2006 International Conference of the IEEE Engineering in Medicine and Biology Society.

[57]  Sreekanth H. Chalasani,et al.  Dissecting a circuit for olfactory behaviour in Caenorhabditis elegans , 2007, Nature.

[58]  Nektarios Tavernarakis,et al.  Nemo: a computational tool for analyzing nematode locomotion , 2007, BMC Neuroscience.

[59]  Diego A. Golombek,et al.  An automated tracking system for Caenorhabditis elegans locomotor behavior and circadian studies application , 2007, Journal of Neuroscience Methods.

[60]  Wenhui Wang,et al.  A Micropositioning System with Real-Time Feature Extraction Capability for Quantifying C. elegans Locomotive Behavior , 2007, 2007 IEEE International Conference on Automation Science and Engineering.

[61]  Laurent Ségalat,et al.  Invertebrate animal models of diseases as screening tools in drug discovery. , 2007, ACS chemical biology.

[62]  Paul W Sternberg,et al.  Epidermal growth factor signaling induces behavioral quiescence in Caenorhabditis elegans , 2007, Nature Neuroscience.

[63]  Pamela C. Cosman,et al.  AUTOMATED TRACKING OF MULTIPLE C. ELEGANS WITH ARTICULATED MODELS , 2007, 2007 4th IEEE International Symposium on Biomedical Imaging: From Nano to Macro.

[64]  Nicolas Roussel,et al.  A Computational Model for C. elegans Locomotory Behavior: Application to Multiworm Tracking , 2007, IEEE Transactions on Biomedical Engineering.

[65]  Paula Ribeiro,et al.  The serotonin receptor SER‐1 (5HT2ce) contributes to the regulation of locomotion in Caenorhabditis elegans , 2007, Developmental neurobiology.