Agonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions.

Pentameric ligand-gated ion channels (pLGICs) activated by the inhibitory neurotransmitter γ-aminobutyric acid (GABA) are expressed widely in both vertebrate and invertebrate species. One of the best characterised insect GABA-gated chloride channels is RDL, an abbreviation of 'resistance to die...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Charles L C Smelt, Victoria R Sanders, Alin M Puinean, Stuart J Lansdell, Jim Goodchild, Neil S Millar
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/408c4c3db8234580a3db059b44035dff
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:408c4c3db8234580a3db059b44035dff
record_format	dspace
spelling	oai:doaj.org-article:408c4c3db8234580a3db059b44035dff2021-12-02T20:15:35ZAgonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions.1932-620310.1371/journal.pone.0254251https://doaj.org/article/408c4c3db8234580a3db059b44035dff2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0254251https://doaj.org/toc/1932-6203Pentameric ligand-gated ion channels (pLGICs) activated by the inhibitory neurotransmitter γ-aminobutyric acid (GABA) are expressed widely in both vertebrate and invertebrate species. One of the best characterised insect GABA-gated chloride channels is RDL, an abbreviation of 'resistance to dieldrin', that was originally identified by genetic screening in Drosophila melanogaster. Here we have cloned the analogous gene from the bumblebee Bombus terrestris audax (BtRDL) and examined its pharmacological properties by functional expression in Xenopus oocytes. Somewhat unexpectedly, the sensitivity of BtRDL to GABA, as measured by its apparent affinity (EC50), was influenced by heterologous expression conditions. This phenomenon was observed in response to alterations in the amount of cRNA injected; the length of time that oocytes were incubated before functional analysis; and by the presence or absence of a 3' untranslated region. In contrast, similar changes in expression conditions were not associated with changes in apparent affinity with RDL cloned from D. melanogaster (DmRDL). Changes in apparent affinity with BtRDL were also observed following co-expression of a chaperone protein (NACHO). Similar changes in apparent affinity were observed with an allosteric agonist (propofol) and a non-competitive antagonist (picrotoxinin), indicating that expression-depended changes are not restricted to the orthosteric agonist binding site. Interestingly, instances of expression-dependent changes in apparent affinity have been reported previously for vertebrate glycine receptors, which are also members of the pLGIC super-family. Our observations with BtRDL are consistent with previous data obtained with vertebrate glycine receptors and indicates that agonist and antagonist apparent affinity can be influenced by the level of functional expression in a variety of pLGICs.Charles L C SmeltVictoria R SandersAlin M PuineanStuart J LansdellJim GoodchildNeil S MillarPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 7, p e0254251 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Charles L C Smelt Victoria R Sanders Alin M Puinean Stuart J Lansdell Jim Goodchild Neil S Millar Agonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions.
description	Pentameric ligand-gated ion channels (pLGICs) activated by the inhibitory neurotransmitter γ-aminobutyric acid (GABA) are expressed widely in both vertebrate and invertebrate species. One of the best characterised insect GABA-gated chloride channels is RDL, an abbreviation of 'resistance to dieldrin', that was originally identified by genetic screening in Drosophila melanogaster. Here we have cloned the analogous gene from the bumblebee Bombus terrestris audax (BtRDL) and examined its pharmacological properties by functional expression in Xenopus oocytes. Somewhat unexpectedly, the sensitivity of BtRDL to GABA, as measured by its apparent affinity (EC50), was influenced by heterologous expression conditions. This phenomenon was observed in response to alterations in the amount of cRNA injected; the length of time that oocytes were incubated before functional analysis; and by the presence or absence of a 3' untranslated region. In contrast, similar changes in expression conditions were not associated with changes in apparent affinity with RDL cloned from D. melanogaster (DmRDL). Changes in apparent affinity with BtRDL were also observed following co-expression of a chaperone protein (NACHO). Similar changes in apparent affinity were observed with an allosteric agonist (propofol) and a non-competitive antagonist (picrotoxinin), indicating that expression-depended changes are not restricted to the orthosteric agonist binding site. Interestingly, instances of expression-dependent changes in apparent affinity have been reported previously for vertebrate glycine receptors, which are also members of the pLGIC super-family. Our observations with BtRDL are consistent with previous data obtained with vertebrate glycine receptors and indicates that agonist and antagonist apparent affinity can be influenced by the level of functional expression in a variety of pLGICs.
format	article
author	Charles L C Smelt Victoria R Sanders Alin M Puinean Stuart J Lansdell Jim Goodchild Neil S Millar
author_facet	Charles L C Smelt Victoria R Sanders Alin M Puinean Stuart J Lansdell Jim Goodchild Neil S Millar
author_sort	Charles L C Smelt
title	Agonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions.
title_short	Agonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions.
title_full	Agonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions.
title_fullStr	Agonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions.
title_full_unstemmed	Agonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions.
title_sort	agonist and antagonist properties of an insect gaba-gated chloride channel (rdl) are influenced by heterologous expression conditions.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/408c4c3db8234580a3db059b44035dff
work_keys_str_mv	AT charleslcsmelt agonistandantagonistpropertiesofaninsectgabagatedchloridechannelrdlareinfluencedbyheterologousexpressionconditions AT victoriarsanders agonistandantagonistpropertiesofaninsectgabagatedchloridechannelrdlareinfluencedbyheterologousexpressionconditions AT alinmpuinean agonistandantagonistpropertiesofaninsectgabagatedchloridechannelrdlareinfluencedbyheterologousexpressionconditions AT stuartjlansdell agonistandantagonistpropertiesofaninsectgabagatedchloridechannelrdlareinfluencedbyheterologousexpressionconditions AT jimgoodchild agonistandantagonistpropertiesofaninsectgabagatedchloridechannelrdlareinfluencedbyheterologousexpressionconditions AT neilsmillar agonistandantagonistpropertiesofaninsectgabagatedchloridechannelrdlareinfluencedbyheterologousexpressionconditions
_version_	1718374521043419136

Agonist and antagonist properties of an insect GABA-gated chloride channel (RDL) are influenced by heterologous expression conditions.

Ejemplares similares