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PharmacoGenetic targeting of a C. elegans essential neuron provides an in vivo screening for novel modulators of nematode ion channel function

PharmacoGenetic targeting of a C. elegans essential neuron provides an in vivo screening for novel modulators of nematode ion channel function
PharmacoGenetic targeting of a C. elegans essential neuron provides an in vivo screening for novel modulators of nematode ion channel function
Chemical or drug treatments are successfully used to treat parasitic nematode infections that impact human, animal and plant health. Many of these exert their effects through modifying neural function underpinning behaviours essential for parasite viability. Selectivity against the parasite may be achieved through distinct pharmacological properties of the parasite nervous system, as exemplified by the success of the ivermectin which target a glutamate-gated chloride channel found only in invertebrates. Despite the success of the ivermectins, emerging resistance and concerns around eco-toxicity are driving the search for new nematocidal chemicals or drugs. Here, we describe the potential of a 5-HT-gated chloride channel MOD-1, which is involved in vital parasite behaviours with constrained distribution in the invertebrate phyla. This ion channel has potential pharmacophores that could be targeted by new nematocidal chemicals and drugs. We have developed a microtiter based bioassay for MOD-1 pharmacology based on its ectopic expression in the Caenorhabditis elegans essential neuron M4. We have termed this technology ‘PhaGeM4’ for ‘Pharmacogenetic targeting of M4 neuron’. Exposure of transgenic worms harbouring ectopically expressed MOD-1 to 5-HT results in developmental arrest. By additional expression of a fluorescence marker in body wall muscle to monitor growth we demonstrate that this assay is suitable for the identification of receptor agonists and antagonists. Indeed, the developmental progression is a robustly quantifiable bioassay that resolves MOD-1 activation by quipazine, 5-carboxyamidotryptamine and fluoxetine and highlight methiothepin as a potent antagonist. This assay has the intrinsic ability to highlight compounds with optimal bioavailability and furthermore to filter out off-target effects. It can be extended to the investigation of other classes of membrane receptors and modulators of neuronal excitation. This approach based on heterologous modulation of the essential M4 neuron function offers a route to discover new effective and selective anthelmintics potentially less confounded by disruptive environmental impact.
5-HT, Cys-loop receptors, Ligand-gated chloride channel, MOD-1, Monoxenic culture, Pharyngeal function, Quipazine
0048-3575
Calahorro, Fernando
81b5dd15-c2cc-4470-98d3-9952aca6fb64
Chapman, Mark
8bac4a92-bfa7-4c3c-af29-9af852ef6383
Dudkiewicz, Katarzyna
e7b14c3c-fefa-4a76-afda-b803187a6b4a
Holden-dye, Lindy
8032bf60-5db6-40cb-b71c-ddda9d212c8e
O'connor, Vincent
8021b06c-01a0-4925-9dde-a61c8fe278ca
Calahorro, Fernando
81b5dd15-c2cc-4470-98d3-9952aca6fb64
Chapman, Mark
8bac4a92-bfa7-4c3c-af29-9af852ef6383
Dudkiewicz, Katarzyna
e7b14c3c-fefa-4a76-afda-b803187a6b4a
Holden-dye, Lindy
8032bf60-5db6-40cb-b71c-ddda9d212c8e
O'connor, Vincent
8021b06c-01a0-4925-9dde-a61c8fe278ca

Calahorro, Fernando, Chapman, Mark, Dudkiewicz, Katarzyna, Holden-dye, Lindy and O'connor, Vincent (2022) PharmacoGenetic targeting of a C. elegans essential neuron provides an in vivo screening for novel modulators of nematode ion channel function. Pesticide Biochemistry and Physiology, 186, [105152]. (doi:10.1016/j.pestbp.2022.105152).

Record type: Article

Abstract

Chemical or drug treatments are successfully used to treat parasitic nematode infections that impact human, animal and plant health. Many of these exert their effects through modifying neural function underpinning behaviours essential for parasite viability. Selectivity against the parasite may be achieved through distinct pharmacological properties of the parasite nervous system, as exemplified by the success of the ivermectin which target a glutamate-gated chloride channel found only in invertebrates. Despite the success of the ivermectins, emerging resistance and concerns around eco-toxicity are driving the search for new nematocidal chemicals or drugs. Here, we describe the potential of a 5-HT-gated chloride channel MOD-1, which is involved in vital parasite behaviours with constrained distribution in the invertebrate phyla. This ion channel has potential pharmacophores that could be targeted by new nematocidal chemicals and drugs. We have developed a microtiter based bioassay for MOD-1 pharmacology based on its ectopic expression in the Caenorhabditis elegans essential neuron M4. We have termed this technology ‘PhaGeM4’ for ‘Pharmacogenetic targeting of M4 neuron’. Exposure of transgenic worms harbouring ectopically expressed MOD-1 to 5-HT results in developmental arrest. By additional expression of a fluorescence marker in body wall muscle to monitor growth we demonstrate that this assay is suitable for the identification of receptor agonists and antagonists. Indeed, the developmental progression is a robustly quantifiable bioassay that resolves MOD-1 activation by quipazine, 5-carboxyamidotryptamine and fluoxetine and highlight methiothepin as a potent antagonist. This assay has the intrinsic ability to highlight compounds with optimal bioavailability and furthermore to filter out off-target effects. It can be extended to the investigation of other classes of membrane receptors and modulators of neuronal excitation. This approach based on heterologous modulation of the essential M4 neuron function offers a route to discover new effective and selective anthelmintics potentially less confounded by disruptive environmental impact.

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Accepted/In Press date: 16 June 2022
e-pub ahead of print date: 20 June 2022
Published date: 1 August 2022
Keywords: 5-HT, Cys-loop receptors, Ligand-gated chloride channel, MOD-1, Monoxenic culture, Pharyngeal function, Quipazine

Identifiers

Local EPrints ID: 457997
URI: http://eprints.soton.ac.uk/id/eprint/457997
ISSN: 0048-3575
PURE UUID: 91ca802b-fe80-4754-a403-815744fe0226
ORCID for Mark Chapman: ORCID iD orcid.org/0000-0002-7151-723X
ORCID for Lindy Holden-dye: ORCID iD orcid.org/0000-0002-9704-1217
ORCID for Vincent O'connor: ORCID iD orcid.org/0000-0003-3185-5709

Catalogue record

Date deposited: 23 Jun 2022 18:29
Last modified: 24 Jul 2022 01:31

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Contributors

Author: Fernando Calahorro
Author: Mark Chapman ORCID iD
Author: Katarzyna Dudkiewicz

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