Droplet microfluidics for characterizing the neurotoxin-induced responses in individual Caenorhabditis elegans.

A droplet-based microfluidic device integrated with a novel floatage-based trap array and a tapered immobilization channel array was presented for characterizing the neurotoxin-induced multiple responses in individual Caenorhabditis elegans (C. elegans) continuously. The established device enabled the evaluations of movement and fluorescence imaging analysis of individual C. elegans simultaneously. The utility of this device was demonstrated by the pharmacological evaluation of neurotoxin (6-hydroxydopamine, 6-OHDA) triggered mobility defects, neuron degeneration and oxidative stress in individual worms. Exposure of living worms to 6-OHDA could cause obvious mobility defects, selective degeneration of dopaminergic (DAergic) neurons, and increased oxidative stress in a dose dependent manner. These results are important towards the understanding of mechanisms leading to DAergic toxicity by neurotoxin and will be of benefit for the screening of new therapeutics for neurodegenerative diseases. This device was simple, stable and easy to operate, with the potential to facilitate whole-animal assays and drug screening in a high throughput manner at single animal resolution.

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

[2]  J. Burgess,et al.  Genetics of lifespan in C. elegans: molecular diversity, physiological complexity, mechanistic simplicity. , 2001, Trends in genetics : TIG.

[3]  Bingcheng Lin,et al.  A programmable microvalve-based microfluidic array for characterization of neurotoxin-induced responses of individual C. elegans. , 2009, Biomicrofluidics.

[4]  Bingcheng Lin,et al.  Droplet-based microfluidic system for individual Caenorhabditis elegans assay. , 2008, Lab on a chip.

[5]  William R Schafer,et al.  Deciphering the Neural and Molecular Mechanisms of C. elegans Behavior , 2005, Current Biology.

[6]  C. Nichols,et al.  Identification of neuroprotective compounds of Caenorhabditis elegans dopaminergic neurons against 6-OHDA , 2007, Journal of Molecular Neuroscience.

[7]  Adela Ben-Yakar,et al.  CO2 and compressive immobilization of C. elegans on-chip. , 2009, Lab on a chip.

[8]  David H. Hall,et al.  Neurotoxin-induced degeneration of dopamine neurons in Caenorhabditis elegans , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[9]  Adela Ben-Yakar,et al.  Femtosecond laser nanoaxotomy lab-on-a-chip for in vivo nerve regeneration studies , 2008, Nature Methods.

[10]  Jim Zoval,et al.  Automated microfluidic compact disc (CD) cultivation system of Caenorhabditis elegans , 2007 .

[11]  C. Behl,et al.  Novel imine antioxidants at low nanomolar concentrations protect dopaminergic cells from oxidative neurotoxicity , 2009, Journal of neurochemistry.

[12]  R. Blakely,et al.  C. elegans: a novel pharmacogenetic model to study Parkinson's disease. , 2001, Parkinsonism & related disorders.

[13]  G. Whitesides,et al.  Fabrication of microfluidic systems in poly(dimethylsiloxane) , 2000, Electrophoresis.

[14]  Min Han,et al.  Control and integration of cell signaling pathways during C. Elegans vulval development , 1996, BioEssays : news and reviews in molecular, cellular and developmental biology.

[15]  Mehmet Fatih Yanik,et al.  Microfluidic system for on-chip high-throughput whole-animal sorting and screening at subcellular resolution , 2007, Proceedings of the National Academy of Sciences.

[16]  K. Caldwell,et al.  Acetaminophen attenuates dopamine neuron degeneration in animal models of Parkinson's disease , 2008, Neuroscience Letters.

[17]  G. Whitesides,et al.  Generation of monodisperse particles by using microfluidics: control over size, shape, and composition. , 2005, Angewandte Chemie.

[18]  Daniel T. Chiu,et al.  Single-synapse ablation and long-term imaging in live C. elegans , 2008, Journal of Neuroscience Methods.

[19]  Matthew M. Crane,et al.  Automated on-chip rapid microscopy, phenotyping and sorting of C. elegans , 2008, Nature Methods.

[20]  Michael Brand,et al.  Micro fluid segment technique for screening and development studies on Danio rerio embryos. , 2007, Lab on a chip.

[21]  Cori Bargmann,et al.  Microfluidics for in vivo imaging of neuronal and behavioral activity in Caenorhabditis elegans , 2007, Nature Methods.

[22]  P. Riederer,et al.  Caenorhabditis elegans MPP+ Model of Parkinson’s Disease for High-Throughput Drug Screenings , 2004, Neurodegenerative Diseases.

[23]  P. Jenner,et al.  Oxidative stress in Parkinson's disease , 2003, Annals of neurology.

[24]  George Perry,et al.  Oxidative stress and neurotoxicity. , 2008, Chemical research in toxicology.

[25]  Kwanghun Chung,et al.  Computer-enhanced high-throughput genetic screens of C. elegans in a microfluidic system. , 2009, Lab on a chip.

[26]  R. Baumeister,et al.  Caenorhabditis elegans as a Model System for Parkinson’s Disease , 2007, Neurodegenerative Diseases.

[27]  Mehmet Fatih Yanik,et al.  Sub-cellular precision on-chip small-animal immobilization, multi-photon imaging and femtosecond-laser manipulation. , 2008, Lab on a chip.

[28]  J. Apfeld,et al.  A microfabricated array of clamps for immobilizing and imaging C. elegans. , 2007, Lab on a chip.

[29]  D. Chiu,et al.  Selective encapsulation of single cells and subcellular organelles into picoliter- and femtoliter-volume droplets. , 2005, Analytical chemistry.

[30]  Cornelia I. Bargmann,et al.  Oxygen sensation and social feeding mediated by a C. elegans guanylate cyclase homologue , 2004, Nature.

[31]  K. Lowe,et al.  Perfluorochemicals: their applications and benefits to cell culture. , 1998, Trends in biotechnology.

[32]  Monpichar Srisa-Art,et al.  High-throughput DNA droplet assays using picoliter reactor volumes. , 2007, Analytical chemistry.

[33]  J. Qin,et al.  Maze exploration and learning in C. elegans. , 2007, Lab on a chip.

[34]  J. S. Johnson,et al.  Biocompatible surfactants for water-in-fluorocarbon emulsions. , 2008, Lab on a chip.

[35]  Helen Song,et al.  Reactions in droplets in microfluidic channels. , 2006, Angewandte Chemie.

[36]  Andreas Schober,et al.  Classic toxin-induced animal models of Parkinson’s disease: 6-OHDA and MPTP , 2004, Cell and Tissue Research.

[37]  Claudia M Testa,et al.  Rotenone induces oxidative stress and dopaminergic neuron damage in organotypic substantia nigra cultures. , 2005, Brain research. Molecular brain research.