Functional characterisation of a nicotinic acetylcholine receptor α subunit from the brown dog tick, Rhipicephalus sanguineus

Lees, Kristin; Jones, Andrew K.; Matsuda, Kazuhiko; Akamatsu, Miki; Sattelle, David B.; Woods, Debra J.; Bowman, Alan K.

doi:10.1016/j.ijpara.2013.11.002

Cited by 16 publications

(10 citation statements)

References 66 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Closely related proteins include sequences from the wolf spider Pardosa pseudoannulata and the tick Rhipicephalus sanguineus that share 67% and 63% similarity with CsnAChα ( Fig 5 ). Both of these sequences have been expressed in Xenopus oocytes, but similarly to most invertebrate nACh subunits, they failed to function as homomultimers [ 8 , 70 ]. When co-expressed with nAChβ subunits, they formed functional ACh and nicotine sensitive receptors.…”

Section: Resultsmentioning

confidence: 99%

Transcriptome Analysis of the Central and Peripheral Nervous Systems of the Spider Cupiennius salei Reveals Multiple Putative Cys-Loop Ligand Gated Ion Channel Subunits and an Acetylcholine Binding Protein

2015

View full text Add to dashboard Cite

Invertebrates possess a diverse collection of pentameric Cys-loop ligand gated ion channel (LGIC) receptors whose molecular structures, evolution and relationships to mammalian counterparts have been intensely investigated in several clinically and agriculturally important species. These receptors are targets for a variety of control agents that may also harm beneficial species. However, little is known about Cys-loop receptors in spiders, which are important natural predators of insects. We assembled de novo transcriptomes from the central and peripheral nervous systems of the Central American wandering spider Cupiennius salei, a model species for neurophysiological, behavioral and developmental studies. We found 15 Cys-loop receptor subunits that are expected to form anion or cation permeable channels, plus a putative acetylcholine binding protein (AChBP) that has only previously been reported in molluscs and one annelid. We used phylogenetic and sequence analysis to compare the spider subunits to homologous receptors in other species and predicted the 3D structures of each protein using the I-Tasser server. The quality of homology models improved with increasing sequence identity to the available high-resolution templates. We found that C. salei has orthologous γ-aminobutyric acid (GABA), GluCl, pHCl, HisCl and nAChα LGIC subunits to other arthropods, but some subgroups are specific to arachnids, or only to spiders. C. salei sequences were phylogenetically closest to gene fragments from the social spider, Stegodyphus mimosarum, indicating high conservation within the Araneomorphae suborder of spiders. C. salei sequences had similar ligand binding and transmembrane regions to other invertebrate and vertebrate LGICs. They also had motifs associated with high sensitivity to insecticides and antiparasitic agents such as fipronil, dieldrin and ivermectin. Development of truly selective control agents for pest species will require information about the molecular structure and pharmacology of Cys-loop receptors in beneficial species.

show abstract

Section: Resultsmentioning

confidence: 99%

Transcriptome Analysis of the Central and Peripheral Nervous Systems of the Spider Cupiennius salei Reveals Multiple Putative Cys-Loop Ligand Gated Ion Channel Subunits and an Acetylcholine Binding Protein

2015

View full text Add to dashboard Cite

show abstract

“…Mammalian Cys‐loop receptors include anion‐selective γ‐aminobutyric acid type A (GABA A ) and glycine receptors, and cation‐selective nicotinic acetylcholine (nACh) and 5‐HT 3 receptors (Miller and Smart, ). Invertebrates have several types of GABA A and nACh receptors as well as glutamate‐ (GluCl), histamine‐ (HisCl), and pH‐ (pHCl) gated Cl channels (Hosie et al, ; Ozoe et al, ; Lees et al, ). Invertebrate Cys‐loop receptors are prime targets for widely used control agents against agricultural pests and disease vectors (Tomizawa and Casida, ; Wolstenholme, ).…”

mentioning

confidence: 99%

Expression of Cys‐loop receptor subunits and acetylcholine binding protein in the mechanosensory neurons, glial cells, and muscle tissue of the spider Cupiennius salei

Liu

French

Torkkeli

2016

J of Comparative Neurology

View full text Add to dashboard Cite

The central and peripheral nervous system transcriptomes of the spider Cupiennius salei have 15 Cys-loop receptor subunits and an acetylcholine-binding protein (AChBP). Twelve subunits are predicted to form anion channels gated by γ-aminobutyric acid (GABA), glutamate, histamine, or changes in pH, and three are putative ACh-gated cation channels. Spiders have a variety of mechanosensilla and proprioceptive organs that are innervated by efferents in their peripherally located parts, and efferents also innervate muscle fibers. We investigated Cys-loop gene expression in muscle tissue by qPCR and localized this expression in mechanosensilla via in situ hybridization. The cuticular mechanosensory neurons had only CsGABArdl and CspHCl2 subunits, whereas the muscle tissue expressed a wider variety of subunits, especially CsGABAgrd, CsGABA β, CsGluCl1 and CspHCl, but very low levels of the CsGABArdl or CsnACh subunits. An nACh non-α subunit was expressed in a group of unidentified cells in the hypodermis and at low level in the muscle tissue, but the physiological function of this subunit is unknown. The CsnAChα subunit was not expressed in sensory neurons and was expressed at extremely low level in the muscle tissue. None of the probes gave signals in proprioceptive joint receptors, suggesting that efferent innervation to this sense organ employs other receptor types. CsAChBP and a glia-specific homeodomain CsREPO were both expressed in glial cells that surround sensory neurons and also in muscle tissue, probably around the nerve endings of the neuromuscular junction. These locations have large numbers of synapses, suggesting that AChBP may have a function in modulating synaptic transmission. J. Comp. Neurol. 525:1139-1154, 2017. © 2016 Wiley Periodicals, Inc.

show abstract

“…Using R. sanguineus nAChR α1 subunit, it was found that its expression into X. laevis oocytes failed to function as a homomer. When co-expressed with chicken β2 subunit, the chimeric α1β2 nAChR evoked inward currents with a slow activation and deactivation kinetics, in response to ACh and nicotine ( Lees et al, 2014 ). We however argued that the functional properties of expressed receptors could be influenced by the host heterologous cell expression system, in terms of activation and deactivation currents induced by agonists.…”

Section: Discussionmentioning

confidence: 99%

“…Transcriptomic analyses on the tick Rhipicephalus sanguineus and Dermacentor variabilis synganglia have shown that they expressed cDNAs encoding for nAChR subunits ( Lees et al, 2010 ; Bissinger et al, 2011 ). Recently, Lees et al, demonstrated that R. sanguineus α1 subunit co-expressed with the chicken β2 subunit, in the Xenopus oocytes , evoked acetylcholine (ACh) and nicotine currents, but the expressed hybrid receptor was insensitive to IMI ( Lees et al, 2014 ). We propose that different nAChR subtypes exist in the tick nervous system and account for the binding of neonicotinoid insecticides.…”

Section: Introductionmentioning

confidence: 99%

Nicotinic acetylcholine receptors in the synganglion of the tick Ixodes ricinus: Functional characterization using membrane microtransplantation

Mauff

Chouikh²,

Cartereau³

et al. 2020

International Journal for Parasitology: Drugs and Drug Resistan

View full text Add to dashboard Cite

Nicotinic acetylcholine receptors are an important class of excitatory receptors in the central nervous system of arthropods. In the ticks Ixodes ricinus , the functional and pharmacological properties of nicotinic receptors located in their neurons are still unknown. The objective of this study was to characterize the pharmacological properties of tick nicotinic receptors using membrane microtransplantation in Xenopus laevis oocytes and two-electrodes voltage clamp method. The membranes microtransplanted were extracted from the tick synganglion. We found that oocytes microtransplanted with tick synganglion membranes expressed nicotinic acetylcholine receptor subtypes which were activated by acetylcholine (1 mM) and nicotine (1 mM). Currents induced by pressure application of acetylcholine and nicotine were diminished by 10 nM α-bungarotoxin and methyllycaconitine, suggesting that they expressed two subtypes of nicotinic receptors, α-bungarotoxin-sensitive and -insensitive, respectively. In addition, we found that nicotine receptors expressed in the synganglion membranes were poorly sensitive to the neonicotinoid insecticides clothianidin (CLT), imidacloprid (IMI), acetamiprid (ACE) and thiamethoxam (TMX), in agreement with their lack of activity as acaricides. Interestingly, current amplitudes were strongly potentialized in the presence of 1 μM PNU-120596. CLT was more active as an agonist than IMI, TMX and ACE. Finally, we demonstrated that microtransplantation of purified membrane from the tick synganglion can be a valuable tool for the development and screening of compounds targeting tick nicotinic acetylcholine receptor subtypes.

show abstract

Functional characterisation of a nicotinic acetylcholine receptor α subunit from the brown dog tick, Rhipicephalus sanguineus

Cited by 16 publications

References 66 publications

Transcriptome Analysis of the Central and Peripheral Nervous Systems of the Spider Cupiennius salei Reveals Multiple Putative Cys-Loop Ligand Gated Ion Channel Subunits and an Acetylcholine Binding Protein

Transcriptome Analysis of the Central and Peripheral Nervous Systems of the Spider Cupiennius salei Reveals Multiple Putative Cys-Loop Ligand Gated Ion Channel Subunits and an Acetylcholine Binding Protein

Expression of Cys‐loop receptor subunits and acetylcholine binding protein in the mechanosensory neurons, glial cells, and muscle tissue of the spider Cupiennius salei

Nicotinic acetylcholine receptors in the synganglion of the tick Ixodes ricinus: Functional characterization using membrane microtransplantation

Contact Info

Product

Resources

About