An automated tracking system for Caenorhabditis elegans locomotor behavior and circadian studies application

Automation of simple behavioral patterns, such as locomotor activity, is fundamental for pharmacological and genetic screening studies. Recently, circadian behaviors in locomotor activity and stress responses were reported in the nematode Caenorhabditis elegans, a well-known model in genetics and developmental studies. Here we present a new method for long-term recordings of C. elegans (as well as other similar-sized animals) locomotor activity based on an infrared microbeam scattering. Individual nematodes were cultured in a 96-well microtiter plate; we tested L15, CeMM and E. coli liquid cultures in long-term activity tracking experiments, and found CeMM to be the optimal medium. Treatment with 0.2% azide caused an immediate decrease in locomotor activity as recorded with our system. In addition to the validation of the method (including hardware and software details), we report its application in chronobiological studies. Circadian rhythms in animals entrained to light-dark and constant dark conditions (n=48 and 96 worms, respectively) at 16 degrees C, were analyzed by LS periodograms. We obtained a 24.2+/-0.44 h period (52% of significantly rhythmic animals) in LD, and a 23.1+/-0.40 h period (37.5% of significantly rhythmic animals) under DD. The system is automateable using microcontrollers, of low-cost construction and highly reproducible.

[1]  Giulio Tononi,et al.  Reduced sleep in Drosophila Shaker mutants , 2005, Nature.

[2]  R. Hardeland,et al.  Non‐vertebrate melatonin , 2003, Journal of pineal research.

[3]  T. Johnson,et al.  Production of age-synchronous mass cultures of Caenorhabditis elegans. , 1994, Journal of gerontology.

[4]  P. Clopton,et al.  Seeking tau: A comparison of six methods , 1993 .

[5]  H. Manev,et al.  An automated assay of the behavioral effects of cocaine injections in adult Drosophila , 2004, Journal of Neuroscience Methods.

[6]  H. Manev,et al.  γ-Aminobutyric acid B receptor 1 mediates behavior-impairing actions of alcohol in Drosophila: Adult RNA interference and pharmacological evidence , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[7]  N. Wittenburg,et al.  Thermal avoidance in Caenorhabditis elegans: an approach to the study of nociception. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[8]  A. Tanakadate,et al.  Microcomputerized measurement of the circadian locomotor rhythm in microorganisms , 1985, Physiology & Behavior.

[9]  J. Dunlap,et al.  Analysis of circadian rhythms in Neurospora: overview of assays and genetic and molecular biological manipulation. , 2005, Methods in enzymology.

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

[11]  Harold B. Dowse,et al.  Summing locomotor activity data into “bins”: How to avoid artifact in spectral analysis , 1994 .

[12]  Joel D Levine,et al.  Signal analysis of behavioral and molecular cycles , 2002, BMC Neuroscience.

[13]  Ezio Rosato,et al.  Analysis of locomotor activity rhythms in Drosophila , 2006, Nature Protocols.

[14]  Yueqing Peng,et al.  Long‐term but not short‐term blockade of dopamine release in Drosophila impairs orientation during flight in a visual attention paradigm , 2004, The European journal of neuroscience.

[15]  Jennifer J. Loros,et al.  Chronobiology: Biological Timekeeping , 2009 .

[16]  J. Dunlap,et al.  Eukaryotic circadian systems: cycles in common , 1999, Genes to cells : devoted to molecular & cellular mechanisms.

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

[18]  Kanae Iijima-Ando,et al.  Transgenic cAMP response element reporter flies for monitoring circadian rhythms. , 2005, Methods in enzymology.

[19]  Jonathan Hodgkin,et al.  Introduction to genetics and genomics , 2005 .

[20]  M. Blaxter,et al.  Caenorhabditis elegans has a circadian clock , 2002, Current Biology.

[21]  P. Sassone-Corsi,et al.  Analysis of circadian rhythms in zebrafish. , 2005, Methods in enzymology.

[22]  T. Saigusa,et al.  Circadian Behavioural Rhythm in Caenorhabditis elegans , 2002, Current Biology.

[23]  C. Strayer,et al.  Circadian rhythms in prokaryotes: luciferase as a reporter of circadian gene expression in cyanobacteria. , 1993, Proceedings of the National Academy of Sciences of the United States of America.