Optogenetic analysis of locomotor behaviours in C. elegans using a Worm Behaviour Imaging System
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1
University of Toronto, Undergraduate Program in Neuroscience,Department of Physiology, Faculty of Medicine, Canada
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2
University of Toronto, Undergraduate Program in Engineering Science,Department of Physiology, Faculty of Medicine, Canada
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3
University of Toronto, Department of Physiology, Faculty of Medicine, Canada
Optogenetics is a newly emerging field to optically manipulate cell physiology using genetically encoded molecular tools, as represented by a light-gated cation channel, Channelrhodopsin/ChR2, and light-gated chloride pump, Halorhodopsin/NpHR (Fig.1, ref-1). These novel tools allow us to respectively depolarize or hyperpolarize specific neurons under illumination.
In the Suzuki lab, we are applying the optogenetics in the model animal C.elegans to study the simple nervous system composed of 302 neurons that are individually identified under microscope. Using the transgenic worms expressing ChR2 and NpHR in the neurons of our interest, the excitation light is readily delivered through the transparent body wall in the living and moving animals to “remote control” the activity of the neurons. Thus, we can directly address the role of specific neurons in generating behaviours.
The apparently monotonous crawling locomotion of the worms on agar plate actually contains a rich source of information in addressing the computation of the neuronal circuit that generate behaviours, as revealed by novel approaches such as in vivo neuronal imaging (ref-2). However, worm’s behaviours have conventionally been analyzed by eye observations, and a new technique for quantitative measurement has been desired. Recently, imaging technique has been applied to capture images of moving worms and extract numerous parameters through digital image processing (ref-3). We are further developing “Worm Behaviour Imaging System” to incorporate optogenetics to analyze the locomotor behaviours while “remote controlling” specific worms by ChR2 and NpHR. Transgenic worms expressing NpHR in motor neurons paralyze on yellow-light illumination (ref-1), and we are using the worms to evaluate our system. We are also analyzing transgenic worms expressing ChR2 or NpHR in dopaminergic neurons to address their role in locomotor behaviours.
References
1. Zhang, F., et al. Multimodal fast optical interrogation of neural circuitory. Nature, 446, 633-9, 2007.
2. Suzuki, H., et al. Functional asymmetry in Caenorhabditis elegans taste neurons and its computational role in chemotaxis. Nature, 454, 114-8 (2008).
3. Cronin, J.C., et al. An automated system for measuring parameters of nematode sinusoidal movement. BMC Genetics, 6, 5.
Conference:
B.R.A.I.N. platform in Physiology poster day 2009, Toronto, ON, Canada, 16 Dec - 16 Dec, 2009.
Presentation Type:
Poster Presentation
Topic:
Poster presentations
Citation:
Fu
D,
Wang
G,
Su
P and
Suzuki
H
(2009). Optogenetic analysis of locomotor behaviours in C. elegans using a Worm Behaviour Imaging System.
Front. Neurosci.
Conference Abstract:
B.R.A.I.N. platform in Physiology poster day 2009.
doi: 10.3389/conf.neuro.03.2009.17.013
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Received:
16 Dec 2009;
Published Online:
16 Dec 2009.
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Correspondence:
Donald Fu, University of Toronto, Undergraduate Program in Neuroscience,Department of Physiology, Faculty of Medicine, Toronto, Canada, donald.fu@utoronto.ca