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A comparison of electrically evoked and channel rhodopsin-evoked postsynaptic potentials in the pharyngeal system of Caenorhabditis elegans

A comparison of electrically evoked and channel rhodopsin-evoked postsynaptic potentials in the pharyngeal system of Caenorhabditis elegans
A comparison of electrically evoked and channel rhodopsin-evoked postsynaptic potentials in the pharyngeal system of Caenorhabditis elegans
Dissecting the function of neural circuits requires the capability to stimulate and record from the component neurones. Optimally, the methods employed should enable precise activation of distinct elements within the circuit and high-fidelity readout of the neuronal response. Here we compare two methods for neural stimulation in the pharyngeal system of Caenorhabditis elegans by evoking postsynaptic potentials (PSPs) either by electrical stimulation or by expression of the channelrhodopsin [ChR2(gf)] in cholinergic neurones of the pharyngeal circuit. Using a dissection that isolates the pharynx and its embedded neural system of 20 neurones permits analysis of the neurotransmitter pathways within this microcircuit. We describe protocols for selective electrically evoked or ChR2-mediated cholinergic synaptic events in this circuit. The latter was achieved by generating strains, punc-17::ChR2(gf);yfp, that express ChR2(gf) in cholinergic neurones. PSPs evoked by both electrical and light stimulation exhibited a rapid time-course and were blocked by cholinergic receptor antagonists and rapidly reversed on cessation of the stimulus. Electrically evoked PSPs were also reduced in a hypomorphic mutant for the synaptic vesicle acetylcholine transporter, unc-17, further indicating they are nicotinic cholinergic PSPs. The pharyngeal nervous system is exquisitely sensitive to both electrical and light activation. For the latter, short light pulses of 200 ?s delivered to punc-17::ChR2(gf);yfp are capable of generating full muscle action potentials. We conclude that the application of optogenetic approaches to the C. elegans isolated pharynx preparation opens the way for a precise molecular dissection of synaptic events in the pharyngeal microcircuit by providing a molecular and system level analysis of the synapses that control the feeding behaviour of C. elegans.
electrophysiology, C. elegans, unc-17, acetylcholine, neurotransmitter release, channelrhodopsin, optogenetics
1354-2516
43
Franks, Christopher J.
9842534b-4d3f-4ee8-a07e-3b050f748593
Murray, Caitriona
6fcb874d-75d7-49aa-9219-8f9723d862fe
Ogden, David
44b86358-bb7e-4e01-b5ac-ad52dbb6d0fc
O'Connor, Vincent
8021b06c-01a0-4925-9dde-a61c8fe278ca
Holden-Dye, Lindy
8032bf60-5db6-40cb-b71c-ddda9d212c8e
Franks, Christopher J.
9842534b-4d3f-4ee8-a07e-3b050f748593
Murray, Caitriona
6fcb874d-75d7-49aa-9219-8f9723d862fe
Ogden, David
44b86358-bb7e-4e01-b5ac-ad52dbb6d0fc
O'Connor, Vincent
8021b06c-01a0-4925-9dde-a61c8fe278ca
Holden-Dye, Lindy
8032bf60-5db6-40cb-b71c-ddda9d212c8e

Franks, Christopher J., Murray, Caitriona, Ogden, David, O'Connor, Vincent and Holden-Dye, Lindy (2009) A comparison of electrically evoked and channel rhodopsin-evoked postsynaptic potentials in the pharyngeal system of Caenorhabditis elegans. Invertebrate Neuroscience, 9 (1), 43. (doi:10.1007/s10158-009-0088-8).

Record type: Article

Abstract

Dissecting the function of neural circuits requires the capability to stimulate and record from the component neurones. Optimally, the methods employed should enable precise activation of distinct elements within the circuit and high-fidelity readout of the neuronal response. Here we compare two methods for neural stimulation in the pharyngeal system of Caenorhabditis elegans by evoking postsynaptic potentials (PSPs) either by electrical stimulation or by expression of the channelrhodopsin [ChR2(gf)] in cholinergic neurones of the pharyngeal circuit. Using a dissection that isolates the pharynx and its embedded neural system of 20 neurones permits analysis of the neurotransmitter pathways within this microcircuit. We describe protocols for selective electrically evoked or ChR2-mediated cholinergic synaptic events in this circuit. The latter was achieved by generating strains, punc-17::ChR2(gf);yfp, that express ChR2(gf) in cholinergic neurones. PSPs evoked by both electrical and light stimulation exhibited a rapid time-course and were blocked by cholinergic receptor antagonists and rapidly reversed on cessation of the stimulus. Electrically evoked PSPs were also reduced in a hypomorphic mutant for the synaptic vesicle acetylcholine transporter, unc-17, further indicating they are nicotinic cholinergic PSPs. The pharyngeal nervous system is exquisitely sensitive to both electrical and light activation. For the latter, short light pulses of 200 ?s delivered to punc-17::ChR2(gf);yfp are capable of generating full muscle action potentials. We conclude that the application of optogenetic approaches to the C. elegans isolated pharynx preparation opens the way for a precise molecular dissection of synaptic events in the pharyngeal microcircuit by providing a molecular and system level analysis of the synapses that control the feeding behaviour of C. elegans.

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More information

Published date: March 2009
Keywords: electrophysiology, C. elegans, unc-17, acetylcholine, neurotransmitter release, channelrhodopsin, optogenetics

Identifiers

Local EPrints ID: 142809
URI: http://eprints.soton.ac.uk/id/eprint/142809
ISSN: 1354-2516
PURE UUID: be477f3a-bb38-45d8-8269-c60a9bd6b4ad
ORCID for Christopher J. Franks: ORCID iD orcid.org/0000-0002-5412-7037
ORCID for Vincent O'Connor: ORCID iD orcid.org/0000-0003-3185-5709
ORCID for Lindy Holden-Dye: ORCID iD orcid.org/0000-0002-9704-1217

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Date deposited: 01 Apr 2010 16:07
Last modified: 14 Mar 2024 02:44

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Author: Caitriona Murray
Author: David Ogden

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