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StyletChip: a microfluidic device for recording host invasion behaviour and feeding of plant parasitic nematodes

StyletChip: a microfluidic device for recording host invasion behaviour and feeding of plant parasitic nematodes
StyletChip: a microfluidic device for recording host invasion behaviour and feeding of plant parasitic nematodes
Plant parasitic nematodes (PPNs) infest the roots of crops and cause global losses with a severe economic impact on food production. Current chemical control agents are being removed from use due to environmental and toxicity concerns and there is a need for new approaches to crop protection. A key feature of parasitic behaviour for the majority of PPNs is a hollow stomastyle or odontostyle required for interaction with the host plant and feeding. This lance-like microscopic structure, often called a stylet, protrudes from the mouth of the worm and thrusts in a rhythmic manner to stab the host root. Studying stylet activity presents technical challenges and as a consequence the underlying biology is poorly understood. We have addressed this by designing a microfluidic chip which traps the PPN Globodera pallida and permits the recording of an electrophysiological signal concomitant with stylet thrusting. The PDMS chip incorporates a precisely designed aperture to trap the nematode securely around a mid-point of its body. It is fabricated using a novel combination of conventional photolithography and two photon polymerization. The chip incorporates valves for rapid application of test compounds and integral electrodes to facilitate acquisition of electrical signals. We show that stylet thrusting can be induced by controlled application of 5-HT (serotonin) to the worm. Each thrust and retraction produces an electrical waveform that characterises the physiological activity associated with the worm's behaviour. The ability to reproducibly record the stylet activity of PPNs provides a new platform for nematicide screening that specifically focuses on a behaviour that is integral to the parasite host interaction. This is the first report of a microfluidic chip capable of electrophysiological recording from nematodes other than Caenorhabditis elegans. The unique approach is optimised for trapping and recording from smaller worms or worms with distinct anterior body shapes and may be applied to other species of economic or medical importance.
1473-0197
2447-2455
Hu, Chunxiao
4892b566-6809-42a8-8285-1c1e93aac730
Kearn, James
f10467c2-3498-4f7f-affc-0a528252245d
Urwin, Peter
2d27b7d7-bbcd-4658-b8cb-80836cd9257b
Lilley, Catherine
032c7c73-a779-4784-8686-00d22245be32
O'Connor, Vincent
8021b06c-01a0-4925-9dde-a61c8fe278ca
Holden-Dye, Lindy
8032bf60-5db6-40cb-b71c-ddda9d212c8e
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
Hu, Chunxiao
4892b566-6809-42a8-8285-1c1e93aac730
Kearn, James
f10467c2-3498-4f7f-affc-0a528252245d
Urwin, Peter
2d27b7d7-bbcd-4658-b8cb-80836cd9257b
Lilley, Catherine
032c7c73-a779-4784-8686-00d22245be32
O'Connor, Vincent
8021b06c-01a0-4925-9dde-a61c8fe278ca
Holden-Dye, Lindy
8032bf60-5db6-40cb-b71c-ddda9d212c8e
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174

Hu, Chunxiao, Kearn, James, Urwin, Peter, Lilley, Catherine, O'Connor, Vincent, Holden-Dye, Lindy and Morgan, Hywel (2014) StyletChip: a microfluidic device for recording host invasion behaviour and feeding of plant parasitic nematodes. Lab on a Chip, 14, 2447-2455. (doi:10.1039/C4LC00292J). (PMID:24839944)

Record type: Article

Abstract

Plant parasitic nematodes (PPNs) infest the roots of crops and cause global losses with a severe economic impact on food production. Current chemical control agents are being removed from use due to environmental and toxicity concerns and there is a need for new approaches to crop protection. A key feature of parasitic behaviour for the majority of PPNs is a hollow stomastyle or odontostyle required for interaction with the host plant and feeding. This lance-like microscopic structure, often called a stylet, protrudes from the mouth of the worm and thrusts in a rhythmic manner to stab the host root. Studying stylet activity presents technical challenges and as a consequence the underlying biology is poorly understood. We have addressed this by designing a microfluidic chip which traps the PPN Globodera pallida and permits the recording of an electrophysiological signal concomitant with stylet thrusting. The PDMS chip incorporates a precisely designed aperture to trap the nematode securely around a mid-point of its body. It is fabricated using a novel combination of conventional photolithography and two photon polymerization. The chip incorporates valves for rapid application of test compounds and integral electrodes to facilitate acquisition of electrical signals. We show that stylet thrusting can be induced by controlled application of 5-HT (serotonin) to the worm. Each thrust and retraction produces an electrical waveform that characterises the physiological activity associated with the worm's behaviour. The ability to reproducibly record the stylet activity of PPNs provides a new platform for nematicide screening that specifically focuses on a behaviour that is integral to the parasite host interaction. This is the first report of a microfluidic chip capable of electrophysiological recording from nematodes other than Caenorhabditis elegans. The unique approach is optimised for trapping and recording from smaller worms or worms with distinct anterior body shapes and may be applied to other species of economic or medical importance.

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e-pub ahead of print date: 6 May 2014
Organisations: Electronics & Computer Science

Identifiers

Local EPrints ID: 365413
URI: https://eprints.soton.ac.uk/id/eprint/365413
ISSN: 1473-0197
PURE UUID: 06c4e449-34e0-4bd0-8da0-982eb945d8cc
ORCID for Lindy Holden-Dye: ORCID iD orcid.org/0000-0002-9704-1217
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676

Catalogue record

Date deposited: 04 Jun 2014 13:51
Last modified: 19 Nov 2019 02:04

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Contributors

Author: Chunxiao Hu
Author: James Kearn
Author: Peter Urwin
Author: Catherine Lilley
Author: Hywel Morgan ORCID iD

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