Staff Profiles

Modules taught

• BIOS4004 (Undergrduate) Scientific Skills
• BIOS4005 (Undergrduate) Professional and Experimental Skills
• BIOS5011 (Undergrduate) Molecular Biology (Module leader)
• BIOS6009 (Undergrduate) Independent Studies in Life Sciences
• BIOS6010 (Undergrduate) Project (Module leader)
• BIOS7001 (Masters) Advanced Molecular Techniques (Module leader)
• BIOS7002 (Masters) Genome Science
• BIOS7009 (Masters) Research Project

Supervision

PhDs completed:

Joseph Hawkins, 2015-2019, "The characterisation of bee nicotinic acetylcholine receptors".

Research group membership

REF Group: UoA5: Biomedicine, Cell and Developmental Biology, Ecology and Evolution, Genetics and Genomics

Lab Members

• Dr Andrew Jones, Research Group Leader
• Eleanor Mitchell, PhD Student
• Jay Hands, PhD Student, joint supervision with Dr Casper Breuker

Research grants and awards

BBSRC Oxford Doctoral Training Program iCASE studentship 2018-2022

Oxford Brookes Research Excellence Award 2017

Nigel Groome PhD Studentship 2015-2018

Higher Education Innovation Funding (HEIF), Oxford Brookes University, 2014

The Leverhulme Trust RPG-2012-602: 'Characterising the functional spectrum of the mosquito GABA receptor'.

Research projects

Characterisation of insect cys-loop ligand-gated ion channels including the GABA receptor, RDL, and nicotinic acetylcholine receptors.

Investigating target-site insecticide resistance in mosquitoes from Laos.

Journal articles

• Mitchell EL, Viscarra F, Bermudez I, Hawkins J, Goodchild JA, Jones AK , 'The Apis mellifera alpha 5 nicotinic acetylcholine receptor subunit expresses as a homomeric receptor that is sensitive to serotonin'
  Pesticide Biochemistry and Physiology 182 (2022)
  ISSN: 0048-3575 eISSN: 1095-9939
  Abstract

  Insect nicotinic acetylcholine receptors (nAChRs) are molecular targets of highly effective insecticides such as neonicotinoids. Functional expression of these receptors provides useful insights into their functional and pharmacological properties. Here, we report that the a5 nAChR subunit of the honey bee, Apis mellifera, functionally expresses in Xenopus laevis oocytes, which is the first time a homomeric insect nAChR has been robustly expressed in a heterologous system without the need for chaperone proteins. Using two-electrode voltage-clamp electrophysiology we show that the a5 receptor has low sensitivity to acetylcholine with an EC₅₀ of 2.37 mM. However, serotonin acts as an agonist with a considerably lower EC₅₀ at 119 microM that is also more efficacious than acetylcholine in activating the receptor. Molecular modelling indicates that residues in the complementary binding site may be involved in the selectivity towards serotonin. This is the first report of a ligand-gated ion channel activated by serotonin from an insect and phylogenetic analysis shows that the a5 subunit of A. mellifera and other non-Dipteran insects, including pest species, belong to a distinct subgroup of subunits, which may represent targets for the development of novel classes of insecticides.

  Website 
• Marcombe S, Shimell K, Savage R, Howlett E, Luangamath P, Nilaxay S, Vungkyly V, Baby A, King M, Clarke J, Jeffries C, Jojo J, Lacey E, Bhatty F, Mabika D, Dela Cruz A, Fisher C, Mbadu M, Despiniadis I, Brey PT, Thammavong P and Jones AK, 'Detection of pyrethroid resistance mutations and intron variants in the voltage-gated sodium channel of Aedes (Stegomyia) aegypti and Ae. (Stegomyia) albopictus mosquitoes from Lao People's Democratic Republic'
  Medical and Veterinary Entomology in press (2022)
  ISSN: 0269-283X eISSN: 1365-2915
  Abstract

  In Lao People's Democratic Republic (PDR), Aedes aegypti (Linnaeus 1762) and Ae. albopictus (Skuse 1894) mosquitoes (Diptera: Culicidae) are vectors of arboviral diseases such as dengue. As the treatment for these diseases are limited, control of the vectors with the use of pyrethroid insecticides is still essential. However, mutations in the voltage-gated sodium channel gene (vgsc) giving rise to pyrethroid resistance is threatening vector control programs. Here, we analyzed both Ae. aegypti and Ae. albopictus mosquitoes collected in different districts of Laos (Kaysone Phomvihane, Vangvieng, Saysettha and Xaythany) for vgsc mutations commonly found throughout Asia (S989P, V1016G and F1534C). Sequences of the vgsc gene showed that the F1534C mutation was prevalent in both Aedes species. S989P and V1016G mutations were detected in Ae. aegypti from each site and were always found together. In addition, the mutation T1520I was seen in Ae. albopictus mosquitoes from Saysettha district as well as in all Ae. aegypti samples. Thus, mutations in the vgsc gene of Ae. aegypti are prevalent in the four districts studied indicating growing insecticide resistance throughout Laos. Constant monitoring programmes and alternative strategies for controlling Aedes should be utilized in order to prolong the effectiveness of pyrethroids thereby maximizing vector control.

• Hawkins J, Mitchell EL, Jones AK, 'NACHO permits functional heterologous expression of an insect homomeric a6 nicotinic acetylcholine receptor'
  Pesticide Biochemistry and Physiology 181 (2021)
  ISSN: 0048-3575 eISSN: 1095-9939
  Abstract

  Insect nicotinic acetylcholine receptors (nAChR) are molecular targets of highly effective insecticides. The use of chaperone proteins has been key to successful functional expression of these receptors in heterologous systems, permitting functional and pharmacological studies of insect nAChRs with particular subunit composition. Here, we report the first use of the chaperone protein, NACHO, to enable functional expression of an insect nAChR, the a6 subunit from Apis mellifera, in Xenopus laevis oocytes. This is also the first report of functional expression of a homomeric insect a6 nAChR. Using two-electrode voltage-clamp electrophysiology we show that the acetylcholine EC₅₀ of the a6 receptor is 0.88 microM and that acetylcholine responses are antagonized by a-bungarotoxin. Spinosad showed agonist actions and kept the ion channel open when co-applied with acetylcholine, reinforcing the a6 nAChR subunit as a key molecular target for the spinosyn class of insecticide. The use of NACHO may provide a basis for future expression studies of insect a6 nAChRs, potentially providing a tool for the discovery of novel insecticides.

  Website 
• Jia ZQ, Zhan EL, Zhang SG, Jones AK, Zhu L, Wang YN, Huang QT, Han ZJ, Zhao CQ, 'Sublethal doses of broflanilide prevents molting in the fall armyworm, Spodoptera frugiperda via altering molting hormone biosynthesis'
  Pesticide Biochemistry and Physiology 181 (2021)
  ISSN: 0048-3575 eISSN: 1095-9939
  Abstract

  Broflanilide is a novel insecticide with a unique mode of action on the insect GABA receptor and is registered worldwide for the control of agricultural pests. It shows high efficacy in controlling the fall armyworm (FAW) Spodoptera frugiperda, which is a destructive pest to various crops. FAW were exposed to sublethal concentrations of broflanilide to determine its impact on insect development. Sublethal doses (LD₁₀ and LD₃₀) caused failure of ecdysis, reduced body length of larvae, malformation of pupae, and vestigial wing formation in adults. Also, broflanilide at LD₃₀ significantly reduced the amount of molting hormone (MH) . After exposure to LD₁₀ or LD₃₀ broflanilide, expression of five Halloween genes, which participate in MH biosynthesis, were found to be altered. Specifically, the transcript levels of SfrCYP307A1 (Spook), SfrCYP314A1 (Shade) and SfrCYP315A1 (Shadow) in 3^rd day larvae were significantly decreased as well as SfrCYP302A1 (Disembodied) and SfrCYP306A1 (Phantom) in 5^th day pupae. In contrast, the transcript levels of SfrCYP302A1 in 3^rd day larvae, SfrCYP307A1 and SfrCYP314A1 in 5^th day pupae, and SfrCYP306A1, SfrCYP307A1 and SfrCYP315A1 in 0.5^th day adults were significantly increased. Our results demonstrate that broflanilide caused the failure of ecdysis in FAW possibly by influencing the intake of cholesterol through inhibition of feeding and also via altering expression of genes important for MH biosynthesis.

  Website 
• Huang QT, Sheng CW, Jones AK, Jiang J, Tang T, Han ZJ, Zhao CQ, 'Functional Characteristics of the Lepidopteran Ionotropic GABA Receptor 8916 Subunit Interacting with the LCCH3 or the RDL Subunit '
  Journal of Agricultural and Food Chemistry 69 (39) (2021) pp.11582-11591
  ISSN: 0021-8561 eISSN: 1520-5118
  Abstract

  The inotropic γ-aminobutyric acid (iGABA) receptor is commonly considered as a fast inhibitory channel and is an important insecticide target. Since 1990, RDL, LCCH3, and GRD were successively isolated and found to be potential subunits of the insect iGABA receptor. More recently, one orphan gene named as 8916 was found and considered to be another potential iGABA receptor subunit according to its amino acid sequence. However, little information about 8916 has been reported. Here, the 8916 subunit from Chilo suppressalis was studied to determine whether it can form part of functional iGABA receptors by co-expressing this subunit with CsRDL1 or CsLCCH3 in the Xenopus oocyte system. Cs8916 or CsLCCH3 did not form functional ion channels when expressed alone. However, Cs8916 was able to form heteromeric ion channels when expressed with either CsLCCH3 or CsRDL1. The recombinant heteromeric Cs8916/LCCH3 channel was a cation-selective channel, which was sensitive to GABA or β-alanine. The current of Cs8916/LCCH3 channel was inhibited by dieldrin, endosulfan, fipronil or ethiprole. In contrast, fluralaner, broflanilide and avermectin showed little effect on the Cs8916/LCCH3 channel (IC₅₀s > 10,000 nM). The Cs8916/RDL1 channel was sensitive to GABA, which were significantly different in EC₅₀ and I_max for GABA to homomeric CsRDL1. Fluralaner, fipronil or dieldrin showed antagonistic actions on Cs8916/RDL1. In conclusion, Cs8916 is a potential iGABA receptor subunit, which can interact with CsLCCH3 to generate a cation-selective channel that is sensitive to several insecticides. Also, as Cs8916/RDL1 has a higher EC₅₀ than homomeric CsRDL1, Cs8916 may serve to affect the physiological function of CsRDL1 and therefore play a role in fine-tuning GABAergic signaling.

  Website 
• Jones AK, Goven D, Froger J-A, Bantz A, Raymond V , 'The cys-loop ligand-gated ion channel gene superfamilies of the cockroaches Blattella germanica and Periplaneta americana'
  Pest Management Science 77 (8) (2021) pp.3787-3799
  ISSN: 1526-498X eISSN: 1526-4998
  Abstract

  Background. Cockroaches are serious urban pests that can transfer disease-causing microorganisms as well as trigger allergic reactions and asthma. They are commonly managed by pesticides that act on cys-loop ligand-gated ion channels (cysLGIC). To provide further information that will enhance our understanding of how insecticides act on their molecular targets in cockroaches, we used genome and reverse transcriptase PCR data to characterise the cysLGIC gene superfamilies from Blattella germanica and Periplaneta americana.
  

  Results. The B. germanica and P. americana cysLGIC superfamilies consist of 30 and 32 subunit-encoding genes, respectively, which are the largest insect cysLGIC superfamilies characterized to date. As with other insects, the cockroaches possess ion channels predicted to be gated by acetylcholine, gamma-aminobutyric acid, glutamate and histamine, as well as orthologues of the Drosophila pH-sensitive chloride channel (pHCl), CG8916 and CG12344. The large cysLGIC superfamilies of cockroaches are a result of an expanded number of divergent nicotinic acetylcholine receptor subunits, with B. germanica and P. americana respectively possessing eight and ten subunit genes. Diversity of the cockroach cysLGICs is also broadened by alternative splicing and RNA A-to-I editing. Unusually, both cockroach species possess a second glutamate-gated chloride channel as well as another CG8916 subunit.
  

  Conclusion. These findings on B. germanica and P. americana enhance our understanding of the evolution of the insect cysLGIC superfamily and provide a useful basis for the study of their function, the detection and management of insecticide resistance, and for the development of improved pesticides with greater specificity towards these major pests.
  

  Website 
• Zhong-Qiang J, Yi-Chi Z, Qiu-Tang H, Jones A, Zhao-Jun H, Chun-Qing Z, 'Acute toxicity, bioconcentration, elimination, action mode and detoxification metabolism of broflanilide in zebrafish, Danio rerio'
  Journal of Hazardous Materials 394 (2020)
  ISSN: 0304-3894
  Abstract

  Broflanilide, a novel meta-diamide insecticide, shows high insecticidal activity against agricultural pests and is scheduled to be launched onto the market in 2020. However, little information about its potential toxicological effects on fish has been reported. In this study, broflanilide showed low toxicity to the zebrafish, Danio rerio, with LC50 > 10 mg L-1 at 96 h and also did not inhibit GABA-induced currents of the heteromeric Drα1β2Sγ2 GABA receptor. Broflanilide showed medium bioconcentration level with a bioconcentration factor at steady state (BCFss) of 10.02 and 69.40 in D. rerio at 2.00 mg L-1 and 0.20 mg L-1, respectively. In the elimination process, the concentration of broflanilide rapidly decreased within two days and slowly dropped below the limit of quantification after ten days. In the 2.00 mg L-1 broflanilide treatment, CYP450 activity was significantly increased up to 3.11-fold during eight days. Glutathione-S- transferase (GST) activity significantly increased by 91.44% within four days. In conclusion, the acute toxicity of broflanilide was low, but it might induce chronic toxicity, affecting metabolism. To our knowledge, this is the first report of the toxicological effects of broflanilide on an aquatic organism, which has the potential to guide the use of broflanilide in the field.
  

  Website 
• Marcombe S, Thammavong P, Luangamath P, Chonephetsarath S, Phommavanh N, Lakeomany K, Nilaxay S, Rahmani Z, Saverton PJ, Abdullateef OH, Forward J, Jacob AE, Khadam S, Ali W, Boer C, Kakinuma H, Hawkins J, Longstreeth R, Portwood NM, Smee M, Brown N, Kuyucu NC, Lechmere S, Stieger G, Maithaviphet S, Nambanya S, Brey PT, Jones AK, 'Malaria and Dengue mosquito vectors from Lao PDR show a lack of the rdl mutant allele responsible for cyclodiene insecticide resistance'
  Journal of Medical Entomology 57 (3) (2019) pp.815-823
  ISSN: 0022-2585
  Abstract

  The gamma-aminobutyric acid (GABA) receptor, RDL, plays important roles in neuronal signalling and is the target of highly effective insecticides. A mutation in RDL, commonly A296S, underlies resistance to several insecticides such as cyclodienes. Even though the use of cyclodienes has been banned, the occurrence of mutations substituting A296 is notably high in mosquitoes from several countries. Here we report a survey investigating the prevalence of the Rdl mutant allele in mosquitoes from Laos, a country where mosquito-borne diseases such as malaria and dengue fever are health concerns. Anopheles and Aedes mosquitoes were collected from twelve provinces in Laos. Adult bioassays on Ae. aegypti (Linnaeus) (Diptera: Culicidae) and Ae. albopictus (Skuse) showed that all the populations tested were susceptible to dieldrin (4%) following WHO protocols. Exon 7 from a total of 791 mosquitoes was sequenced to identify the amino acid encoded for at 296 of RDL. Only one of these mosquitoes, Anopheles maculatus rampae (Diptera: Culicidae) from Attapeu, carried the mutant allele being heterozygous for A296S. We therefore found a general lack of the Rdl mutant allele indicating that mosquitoes from Laos are not exposed to insecticides that act on the GABA receptor compared to mosquitoes in several other countries. Identifying the prevalence of the Rdl mutation may help inform the potential use of alternative insecticides that act on the GABA receptor should there be a need to replace pyrethroids in order to prevent/manage resistance.
  

  Website 
• Jones AK, 'How complex can RDL, the insect GABA receptor, get?'
  International Journal of Insect Science 10 (2018)
  ISSN: 1179-5433 eISSN: 1179-5433
  Abstract

  Recently, Taylor-Wells et al. published evidence that the GABA receptor, RDL, from mosquitoes undergo RNA A-to-I editing to generate an extraordinarily large range of isoforms. This editing was found to impact on GABA receptor pharmacology as it influenced the potency of GABA and ivermectin. This highlights RNA editing as a species-specific mechanism to fine tune receptor function as well as possibly increase tolerance of mosquitoes to certain insecticides. This commentary also considers novel findings from analysis of Rdl transcripts from individual mosquitoes taken from different geographical areas.
  

  Website 
• Matthews BJ, Dudchenko O, Kingan S, Koren S, Antoshechkin I, Crawford JE, Glassford WJ, Herre M, Redmond SN, Rose NH, Weedall GD, Wu Y, SS, Brito-Sierra CA, Buckingham SD, Campbell CL, Chan S, Cox E, Evans BR, Fansiri T, Filipović I, Fontaine A, Gloria-Soria A, Hall R, Joardar VS, Jones AK, Kay RGG, Kodali VK, Lee J, Lycett GJ, Mitchell SN, Muehling J, Murphy MR, Omer AD, Partridge FA, Peluso P, Aiden AP, Ramasamy V, Rašić G, Roy S, Saavedra-Rodriguez K, Sharan S, Sharma A, Smith ML, Turner J, Weakley AM, Zhao Z, Akbari OS, Black IV WC, Cao H, Darby AC, Hill CA, Johnston JS, Murphy TD, Raikhel AS, Sattelle DB, Sharakhov IV, White BJ, Zhao L, Aiden EL, Mann RS, Lambrechts L, Powell JR, Sharakhova MV, Tu Z, Robertson HM, McBride CS, Hastie AR, Korlach J, Neafsey DE, Phillippy AM, Vosshall LB, 'Improved reference genome of Aedes aegypti informs arbovirus vector control'
  Nature 563 (7732) (2018) pp.501-507
  ISSN: 0028-0836 eISSN: 1476-4687
  Abstract

  Female Aedes aegypti mosquitoes infect more than 400 million people each year with dangerous viral pathogens including dengue, yellow fever, Zika and chikungunya. Progress in understanding the biology of mosquitoes and developing the tools to fight them has been slowed by the lack of a high-quality genome assembly. Here we combine diverse technologies to produce the markedly improved, fully re-annotated AaegL5 genome assembly, and demonstrate how it accelerates mosquito science. We anchored physical and cytogenetic maps, doubled the number of known chemosensory ionotropic receptors that guide mosquitoes to human hosts and egg-laying sites, provided further insight into the size and composition of the sex-determining M locus, and revealed copy-number variation among glutathione S-transferase genes that are important for insecticide resistance. Using high-resolution quantitative trait locus and population genomic analyses, we mapped new candidates for dengue vector competence and insecticide resistance. AaegL5 will catalyse new biological insights and intervention strategies to fight this deadly disease vector.
  

  Website 
• Jones AK, 'Genomics, cys-loop ligand-gated ion channels and new targets for the control of insect pests and vectors'
  Current Opinion in Insect Science 30 (2018) pp.1-7
  ISSN: 2214-5745
  Abstract Cys-loop ligand-gated ion channels (cysLGICs) play roles in the nervous system. They consist of five subunits arranged around a central ion channel with each subunit being encoded for by a separate gene. In insects, the cysLGIC superfamily commonly consists of 21-25 genes giving rise to several receptor classes such as nicotinic acetylcholine receptors and GABA receptors. Insect cysLGICs are of interest as they are the target of insecticides. Analyses of genome sequences have identified cysLGIC gene superfamilies from different species including crop pests, disease vectors and beneficial insects. This review explores recent studies that have pushed forward our knowledge about this superfamily and considers the potential of developing improved strategies to control insect pests whilst sparing non-target organisms.Website 
• Jones AK, del Villar SG, 'Cloning and functional characterisation of the duplicated RDL subunits from the pea aphid, Acyrthosiphon pisum'
  International Journal of Molecular Sciences 19 (8) (2018)
  ISSN: 1661-6596 eISSN: 1422-0067
  Abstract The insect GABA receptor, RDL (resistance to dieldrin), is a cys-loop ligand-gated ion channel (cysLGIC) that plays central roles in neuronal signaling and is the target of several classes of insecticides. Many insects studied to date possess one Rdl gene, however, there is evidence of two Rdls in aphids. To further characterize this insecticide target from pests that cause millions of dollars’ worth of crop damage each year, we identified the complete cysLGIC gene superfamily of the pea aphid, Acyrthosiphon pisum, using BLAST analysis. This confirmed the presence of two Rdl-like genes (RDL1 and RDL2) that likely arose from a recent gene duplication. When expressed individually in Xenopus laevis oocytes, both subunits formed functional ion channels gated by GABA. Alternative splicing of RDL1 influenced the potency of GABA and the potency of fipronil was different on the RDL1bd splice variant and RDL2. Imidacloprid and clothianidin showed no antagonistic activity on RDL1 whilst 100 M thiacloprid reduced the GABA responses of RDL1 and RDL2 to 55% and 62%, respectively. It is concluded that gene duplication of Rdl may have conferred increased tolerance to natural insecticides and played a role in the evolution of insect cysLGICs. Website 
• Braak N, Neve R, Jones AK, Gibbs M, Breuker CJ, 'The effects of insecticides on butterflies – a review'
  Environmental Pollution 242 (A) (2018) pp.507-518
  ISSN: 0269-7491 eISSN: 1873-6424
  Abstract Pesticides, in particular insecticides, can be very beneficial but have also been found to have harmful side effects on non-target insects. Butterflies play an important role in ecosystems, are well monitored and are recognised as good indicators of environmental health. The amount of information already known about butterfly ecology and the increased availability of genomes make them a very valuable model for the study of non-target effects of pesticide usage. The effects of pesticides are not simply linear, but complex through their interactions with a large variety of biotic and abiotic factors. Furthermore, these effects manifest themselves at a variety of levels, from the molecular to metapopulation level. Research should therefore aim to dissect these complex effects at a number of levels, but as we discuss in this review, this is seldom if ever done in butterflies. We suggest that in order dissect the complex effects of pesticides on butterflies we need to integrate detailed molecular studies, including characterising sequence variability of relevant target genes, with more classical evolutionary ecology; from direct toxicity tests on individual larvae in the laboratory to field studies that consider the potentiation of pesticides by ecologically relevant environmental biotic and abiotic stressors. Such integration would better inform population-level responses across broad geographical scales and provide more in-depth information about the non-target impacts of pesticides.Website 
• Taillebois E, Cartereau A, Jones AK, Thany SH, 'Neonicotinoid insecticides mode of action on insect nicotinic acetylcholine receptors using binding studies'
  Pesticide Biochemistry and Physiology 151 (2018) pp.59-66
  ISSN: 0048-3575 eISSN: 1095-9939
  Abstract Nicotinic acetylcholine receptors (nAChRs) are the main target of neonicotinoid insecticides, which are widely used in crop protection against insect pests. Electrophysiological and molecular approaches have demonstrated the presence of several nAChR subtypes with different affinities for neonicotinoid insecticides. However, the precise mode of action of neonicotinoids on insect nAChRs remains to be elucidated. Radioligand binding studies with [3H]-α-bungarotoxin and [3H]-imidacloprid have proved instructive in understanding ligand binding interactions between insect nAChRs and neonicotinoid insecticides. The precise binding site interactions have been established using membranes from whole body and specific tissues. In this review, we discuss findings concerning the number of nAChR binding sites against neonicotinoid insecticides from radioligand binding studies on native tissues. We summarize the data available in the literature and compare the binding properties of the most commonly used neonicotinoid insecticides in several insect species. Finally, we demonstrate that neonicotinoid-nAChR binding sites are also linked to biological samples used and insect species.Website 
• Taylor-Wells J, Senan A, Bermudez I, Jones AK, 'Species specific RNA A-to-I editing of mosquito RDL modulates GABA potency and influences agonistic, potentiating and antagonistic actions of ivermectin'
  Insect Biochemistry and Molecular Biology 93 (2018) pp.1-11
  ISSN: 0965-1748
  Abstract The insect GABA receptor, RDL, is the target of several classes of pesticides. The peptide sequences of RDL are generally highly conserved between diverse insects. However, RNA A-to-I editing can effectively alter amino acid residues of RDL in a species specific manner, which can affect the potency of GABA and possibly insecticides. We report here that RNA A-to-I editing alters the gene products of Rdl in three mosquito disease vectors, recoding five amino acid residues in RDL of Aedes aegypti and six residues in RDLs of Anopheles gambiae and Culex pipiens, which is the highest extent of editing in RDL observed to date. Analysis of An. gambiae Rdl cDNA sequences identified 24 editing isoforms demonstrating a considerable increase in gene product diversity. RNA editing influenced the potency of the neurotransmitter, GABA, on An. gambiae RDL editing isoforms expressed in Xenopus laevis oocytes, as demonstrated by EC50s ranging from 5 ± 1 to 246 ± 41 M. Fipronil showed similar potency on different editing isoforms, with IC50s ranging from 0.18 ± 0.08 to 0.43 ± 0.09 M. In contrast, editing of An. gambiae RDL affected the activating, potentiating and inhibiting actions of ivermectin. For example, ivermectin potentiated currents induced by GABA at the EC20 concentration in the unedited isoform but not in the fully edited variant. Editing of a residue in the first transmembrane domain or the cys-loop influenced this potentiation, highlighting residues involved in the allosteric mechanisms of cys-loop ligand-gated ion channels. Understanding the interactions of ivermectin with molecular targets may have relevance to mosquito control in areas where people are administered with ivermectin to treat parasitic diseases.Website 
• Taylor-Wells J, Hawkins J, Colombo C, Bermudez I, Jones AK, 'Cloning and functional expression of intracellular loop variants of the honey bee (Apis mellifera) RDL GABA receptor'
  NeuroToxicology 60 (2016) pp.207-213
  ISSN: 0161-813X
  Abstract The insect GABA receptor, RDL (resistance to dieldrin), plays central roles in neuronal signalling and is the target of several classes of insecticides. To study the GABA receptor from an important pollinator species, we cloned Rdl cDNA from the honey bee, Apis mellifera. Three Rdl variants were identified, arising from differential use of splice acceptor sites in the large intracellular loop between transmembrane regions 3 and 4. These variants were renamed from previously, as Amel_RDLvar1, Amel_RDLvar2 and Amel_RDLvar3. When expressed in Xenopus laevis oocytes, the three variants showed no difference in sensitivity to the agonist, GABA, with EC50s of 29 μM, 20 μM and 29 μM respectively. Also, the potencies of the antagonists, fipronil and imidacloprid, were similar on all three variants. Fipronil IC50 values were 0.18 μM, 0.31 μM and 0.20 μM whereas 100 μM imidacloprid reduced the GABA response by 17%, 24% and 31%. The possibility that differential splicing of the RDL intracellular loop may represent a species-specific mechanism leading to insensitivity to insecticides is discussed.Website 
• Sadd B, Barribeau SM, Bloch G, de Graaf DC, Dearden P, Elsik CG, Gadau J, Grimmelikhuijzen CJP, Hasselmann M, Lozier JD, Robertson HM, Smagghe G, Stolle E, Van Vaerenbergh M, Waterhouse RM, Bornberg-Bauer E, Klasberg S, Bennett AK, Câmara F,Guigó R, Hoff K, Mariotti M, Munoz-Torres M, Murphy T, Santesmasses D, Amdam GV, Beckers M, Beye M, Biewer M, Bitondi MMG, Blaxter ML, Bourke AFG, Brown MJF, Buechel SD, Cameron R, Cappelle K, Carolan JC, Christiaens O, Ciborowski KL, Clarke DF, Colgan TJ, David H Collins DH, Cridge AG, DalmayT, Dreier S, du Plessis L, Duncan E, Erler S, Evans J, Falcon T, Flores K, Freitas FCP, Fuchikawa T, Gempe T, Hartfelder K, Hauser F, Helbing S, Humann FC, Irvine F, Jermiin LS, Johnson CE, Johnson RM, Jones AK, Kadowaki T, Kidner JH, Koch V, Köhler A, Kraus FB, Lattorff HMG, Leask M, Lockett GA, Mallon EB, Antonio DSM, Marxer M, Meeus I, Moritz RFA, Nair A, Näpflin K, Nissen I, Niu J, Nunes FMF, Oakeshott JG, Osborne A, Otte M, Pinheiro DG, Rossié N, Rueppell O, Santos CG, Schmid-Hempel R, Schmitt BD, Schulte C, Simões ZLP, Soares MPM, Swevers L, Winnebeck EC, Wolschin F, Yu N, Zdobnov EM, Aqrawi PK, Blankenburg KP, Coyle M, Francisco LF, Hernandez AG, Holder M, Hudson ME, Jackson L, Jayaseelan J, Joshi V, Kovar C, Lee SL, Mata R, Mathew T, Newsham IF, Ngo R, Okwuonu G, Pham C, Pu LL, Saada N, Santibanez J, Simmons D, Thornton R, Venkat A, Walden KKO, Wu YQ, Debyser G, Devreese B, Asher C, Blommaert J, Chipman AD, Chittka L, Fouks B, Liu J, O’Neill MP, Sumner S, Puiu D, Qu J, Salzberg SL, Scherer SE, Muzny DM, Richards S, Robinson GE, Gibbs RA, Schmid-Hempel P, Worley KC, 'The genomes of two key bumblebee species with primitive eusocial organization'
  Genome Biology 16 (2015)
  ISSN: 1474-760X
  Abstract

  Background

  The shift from solitary to social behavior is one of the major evolutionary transitions. Primitively eusocial bumblebees are uniquely placed to illuminate the evolution of highly eusocial insect societies. Bumblebees are also invaluable natural and agricultural pollinators, and there is widespread concern over recent population declines in some species. High-quality genomic data will inform key aspects of bumblebee biology, including susceptibility to implicated population viability threats.

   

  Results

  We report the high quality draft genome sequences of Bombus terrestris and Bombus impatiens, two ecologically dominant bumblebees and widely utilized study species. Comparing these new genomes to those of the highly eusocial honeybee Apis mellifera and other Hymenoptera, we identify deeply conserved similarities, as well as novelties key to the biology of these organisms. Some honeybee genome features thought to underpin advanced eusociality are also present in bumblebees, indicating an earlier evolution in the bee lineage. Xenobiotic detoxification and immune genes are similarly depauperate in bumblebees and honeybees, and multiple categories of genes linked to social organization, including development and behavior, show high conservation. Key differences identified include a bias in bumblebee chemoreception towards gustation from olfaction, and striking differences in microRNAs, potentially responsible for gene regulation underlying social and other traits.

   

  Conclusions

  These two bumblebee genomes provide a foundation for post-genomic research on these key pollinators and insect societies. Overall, gene repertoires suggest that the route to advanced eusociality in bees was mediated by many small changes in many genes and processes, and not by notable expansion or depauperation

   

  Website 
• Taylor-Wells J, Brooke BD, Bermudez I, Jones AK, 'The neonicotinoid imidacloprid, and the pyrethroid deltamethrin, are antagonists of the insect Rdl GABA receptor'
  Journal of Neurochemistry 135 (4) (2015) pp.705-713
  ISSN: 0022-3042 eISSN: 1471-4159
  Abstract

  A mutation in the second transmembrane domain of the GABA receptor subunit, Rdl, is associated with resistance to insecticides such as dieldrin and fipronil. Molecular cloning of Rdl cDNA from a strain of the malaria mosquito, Anopheles gambiae, which is highly resistant to dieldrin revealed this mutation (A296G) as well as another mutation in the third transmembrane domain (T345M). Wild-type, A296G, T345M and A296G + T345M homomultimeric Rdl were expressed in Xenopus laevis oocytes and their sensitivities to fipronil, deltamethrin, 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane (DDT), imidacloprid and spinosad were measured using two-electrode voltage-clamp electrophysiology. Spinosad and DDT had no agonist or antagonist actions on Rdl. However, fipronil, deltamethrin and imidacloprid decreased GABA-evoked currents. These antagonistic actions were either reduced or abolished with the A296G and the A296G + T345M mutations while T345M alone appeared to have no significant effect. In conclusion, this study identifies another mutation in the mosquito Rdl that is associated with insecticide resistance. While T345M itself does not affect insecticide sensitivity, it may serve to offset the structural impact of A296G. The present study also highlights Rdl as a potential secondary target for neonicotinoids and pyrethroids.

  Website 
• Kuzma-Kuzniarska M, Yapp C, Pearson-Jones TW, Jones AK, Hulley PA, 'Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching'
  Journal of Biomedical Optics 19 (1) (2014)
  ISSN: 1083-3668
  Abstract

  Gap junction-mediated intercellular communication influences a variety of cellular activities. In tendons, gap junctions modulate collagen production, are involved in strain-induced cell death, and are involved in the response to mechanical stimulation. The aim of the present study was to investigate gap junction-mediated intercellular communication in healthy human tendon-derived cells using fluorescence recovery after photobleaching (FRAP). The FRAP is a noninvasive technique that allows quantitative measurement of gap junction function in living cells. It is based on diffusion-dependent redistribution of a gap junction-permeable fluorescent dye. Using FRAP, we showed that human tenocytes form functional gap junctions in monolayer and three-dimensional (3-D) collagen I culture. Fluorescently labeled tenocytes following photobleaching rapidly reacquired the fluorescent dye from neighboring cells, while HeLa cells, which do not communicate by gap junctions, remained bleached. Furthermore, both 18 β-glycyrrhetinic acid and carbenoxolone, standard inhibitors of gap junction activity, impaired fluorescence recovery in tendon cells. In both monolayer and 3-D cultures, intercellular communication in isolated cells was significantly decreased when compared with cells forming many cell-to-cell contacts. In this study, we used FRAP as a tool to quantify and experimentally manipulate the function of gap junctions in human tenocytes in both two-dimensional (2-D) and 3-D cultures.

  Website 
• Furutani S, Ihara M, Nishino Y, Akamatsu M, Jones AK, Sattelle DB, Matsuda K, 'Exon 3 splicing and mutagenesis identify residues influencing cell surface density of heterologously-expressed silkworm (Bombyx mori) glutamate-gated chloride channels'
  Molecular Pharmacology 86 (6) (2014) pp.686-695
  ISSN: 0026-895X
  Abstract
  Glutamate-gated chloride channels (GluCls) mediate fast inhibitory neurotransmission in 
  invertebrate nervous systems. I Insect GluCls show alternative splicing and to determine its 
  impact on channel function and pharmacology we isolated GluCl cDNAs from larvae of the 
  silkworm (Bombyx mori). We show that 6 BmGluCl variants are generated by splicing in exons 
  3 and 9 and that exons 3b and 3c are common in the brain and third thoracic ganglion. When 
  expressed in Xenopus laevis oocytes, the three functional exon 3 variants (3a, b, c) all had 
  similar EC50 values for L-glutamate and ivermectin (IVM); however, Imax (the maximum 
  L-glutamate- and IVM-induced response of the channels at saturating concentrations) differed 
  strikingly between variants, with the 3c variant showing the largest L-glutamate- and 
  IVM-induced response. By contrast, a partial deletion detected in exon 9 had a much smaller 
  impact on L-glutamate and IVM actions. Binding assays using [3
  H]IVM indicate that diversity 
  in IVM responses among the GluCl variants are mainly due to the impact on channel assembly, 
  altering receptor cell surface numbers. GluCl variants expressed in HEK293 cells show that 
  structural differences influenced Bmax but not Kd values of [3
  H]IVM. Domain swapping and 
  site-directed mutagenesis identified 4 amino acids in exon 3c as hot spots determining the 
  highest amplitude of the L-glutamate and IVM responses. Modeling the GluCl 3a and 3c 
  variants suggested that 3 of the 4 amino acids contribute to inter-subunit contacts, while the 
  other interacts with the TM2-TM3 linker, influencing the receptor response. 
  Glutamate-gated chloride channels (GluCls) mediate fast inhibitory neurotransmission in invertebrate nervous systems. I Insect GluCls show alternative splicing and to determine its impact on channel function and pharmacology we isolated GluCl cDNAs from larvae of the silkworm (Bombyx mori). We show that 6 BmGluCl variants are generated by splicing in exons 3 and 9 and that exons 3b and 3c are common in the brain and third thoracic ganglion. When expressed in Xenopus laevis oocytes, the three functional exon 3 variants (3a, b, c) all had similar EC50 values for L-glutamate and ivermectin (IVM); however, Imax (the maximum L-glutamate- and IVM-induced response of the channels at saturating concentrations) differed strikingly between variants, with the 3c variant showing the largest L-glutamate- and IVM-induced response. By contrast, a partial deletion detected in exon 9 had a much smaller 
  impact on L-glutamate and IVM actions. Binding assays using [3H]IVM indicate that diversity in IVM responses among the GluCl variants are mainly due to the impact on channel assembly, altering receptor cell surface numbers. GluCl variants expressed in HEK293 cells show that structural differences influenced Bmax but not Kd values of [3H]IVM. Domain swapping and site-directed mutagenesis identified 4 amino acids in exon 3c as hot spots determining the highest amplitude of the L-glutamate and IVM responses. Modeling the GluCl 3a and 3c variants suggested that 3 of the 4 amino acids contribute to inter-subunit contacts, while the other interacts with the TM2-TM3 linker, influencing the receptor response. 
  Website 
• Lees K, Jones AK, Matsuda K, Akamatsu M, Sattelle DB, Woods DJ, Bowman AS, 'Functional characterisation of a nicotinic acetylcholine receptor α subunit from the brown dog tick, Rhipicephalus sanguineus'
  International Journal for Parasitology 44 (1) (2014) pp.75-81
  ISSN: 0020-7519
  Abstract

  Ticks and tick-borne diseases have a major impact on human and animal health worldwide. Current control strategies rely heavily on the use of chemical acaricides, most of which target the CNS and with increasing resistance, new drugs are urgently needed. Nicotinic acetylcholine receptors (nAChRs) are targets of highly successful insecticides. We isolated a full-length nAChR α subunit from a normalised cDNA library from the synganglion (brain) of the brown dog tick, Rhipicephalus sanguineus. Phylogenetic analysis has shown this R. sanguineus nAChR to be most similar to the insect α1 nAChR group and has been named Rsanα1. Rsanα1 is distributed in multiple tick tissues and is present across all life-stages. When expressed in Xenopus laevis oocytes Rsanα1 failed to function as a homomer, with and without the addition of either Caenorhabditis elegans resistance-to-cholinesterase (RIC)-3 or X. laevis RIC-3. When co-expressed with chicken β2 nAChR, Rsanα1 evoked concentration-dependent, inward currents in response to acetylcholine (ACh) and showed sensitivity to nicotine (100 μM) and choline (100 μM). Rsanα1/β2 was insensitive to both imidacloprid (100 μM) and spinosad (100 μM). The unreliable expression of Rsanα1 in vitro suggests that additional subunits or chaperone proteins may be required for more robust expression. This study enhances our understanding of nAChRs in arachnids and may provide a basis for further studies on the interaction of compounds with the tick nAChR as part of a discovery process for novel acaricides.

  Website 
• Scott JG, Warren WC, Beukeboom LW, Bopp D, Clark AG, Giers SD, Hediger M, Jones AK, Kasai S, Leichter CA, Li M, Meisel RP, Minx P, Murphy TD, Nelson DR, Reid WR, Rinkevich FD, Robertson HM, Sackton TB, Sattelle DB, Thibaud-Nissen F, Tomlinson C, van de Zande L, Walden KK, Wilson RK, Liu N, 'Genome of the house fly, Musca domestica L., a global vector of diseases with adaptations to a septic environment'
  Genome Biology 15 (2014)
  ISSN: 1474-760X
  Abstract

  BACKGROUND:

  Adult house flies, Musca domestica L., are mechanical vectors of more than 100 devastating diseases that have severe consequences for human and animal health. House fly larvae play a vital role as decomposers of animal wastes, and thus live in intimate association with many animal pathogens.

  RESULTS:

  We have sequenced and analyzed the genome of the house fly using DNA from female flies. The sequenced genome is 691 Mb. Compared with Drosophila melanogaster, the genome contains a rich resource of shared and novel protein coding genes, a significantly higher amount of repetitive elements, and substantial increases in copy number and diversity of both the recognition and effector components of the immune system, consistent with life in a pathogen-rich environment. There are 146 P450 genes, plus 11 pseudogenes, in M. domestica, representing a significant increase relative to D. melanogaster and suggesting the presence of enhanced detoxification in house flies. Relative to D. melanogaster, M. domestica has also evolved an expanded repertoire of chemoreceptors and odorant binding proteins, many associated with gustation.

  CONCLUSIONS:

  This represents the first genome sequence of an insect that lives in intimate association with abundant animal pathogens. The house fly genome provides a rich resource for enabling work on innovative methods of insect control, for understanding the mechanisms of insecticide resistance, genetic adaptation to high pathogen loads, and for exploring the basic biology of this important pest. The genome of this species will also serve as a close out-group to Drosophila in comparative genomic studies.

   

  Website 
• Bennett HM, Lees K, Harper KM, Jones AK, Sattelle DB, Wonnacott S, Wolstenholme AJ, 'Xenopus laevis RIC-3 enhances the functional expression of the C. elegans homomeric nicotinic receptor, ACR-16, in Xenopus oocytes.'
  Journal of Neurochemistry 123 (6) (2012) pp.911-918
  ISSN: 0022-3042
  Abstract RIC-3 enhances the functional expression of certain nicotinic acetylcholine receptors (nAChRs) in vertebrates and invertebrates and increases the availability of functional receptors in cultured cells and Xenopus laevis oocytes. Maximal activity of RIC-3 may be cell-type dependent, so neither mammalian nor invertebrate proteins is optimal in amphibian oocytes. We cloned the X. laevis ric-3 cDNA and tested the frog protein in oocyte expression studies. X. laevis RIC-3 shares 52% amino acid identity with human RIC-3 and only 17% with that of Caenorhabditis elegans. We used the C. elegans nicotinic receptor, ACR-16, to compare the ability of RIC-3 from three species to enhance receptor expression. In the absence of RIC-3, the proportion of oocytes expressing detectable nAChRs was greatly reduced. Varying the ratio of acr-16 to X. laevis ric-3 cRNAs injected into oocytes had little impact on the total cell current. When X. laevis, human or C. elegans ric-3 cRNAs were co-injected with acr-16 cRNA (1 : 1 ratio), 100 μM acetylcholine induced larger currents in oocytes expressing X. laevis RIC-3 compared with its orthologues. This provides further evidence for a species-specific component of RIC-3 activity, and suggests that X. laevis RIC-3 is useful for enhancing the expression of invertebrate nAChRs in X. laevis oocytes.Website 
• Jones, A K, Rayes D, Al-Diwani A, Maynard, T P R, Jones, R, Hernando G, Buckingham S D, Bouzat C, Sattelle D B, 'A cys-loop mutation in the Caenorhabditis elegans nicotinic receptor subunit UNC-63 impairs but does not abolish channel function'
  Journal of Biological Chemistry 286 (4) (2011) pp.2550-2558
  ISSN: 0021-9258
  Abstract

  The nematode Caenorhabditis elegans is an established model organism for studying neurobiology. UNC-63 is a C. elegans nicotinic acetylcholine receptor (nAChR) α-subunit. It is an essential component of the levamisole-sensitive muscle nAChR (L-nAChR) and therefore plays an important role in cholinergic transmission at the nematode neuromuscular junction. Here, we show that worms with the unc-63(x26) allele, with its αC151Y mutation disrupting the Cys-loop, have deficient muscle function reflected by impaired swimming (thrashing). Single-channel recordings from cultured muscle cells from the mutant strain showed a 100-fold reduced frequency of opening events and shorter channel openings of L-nAChRs compared with those of wild-type worms. Anti-UNC-63 antibody staining in both cultured adult muscle and embryonic cells showed that L-nAChRs were expressed at similar levels in the mutant and wild-type cells, suggesting that the functional changes in the receptor, rather than changes in expression, are the predominant effect of the mutation. The kinetic changes mimic those reported in patients with fast-channel congenital myasthenic syndromes. We show that pyridostigmine bromide and 3,4-diaminopyridine, which are drugs used to treat fast-channel congenital myasthenic syndromes, partially rescued the motility defect seen in unc-63(x26). The C. elegans unc-63(x26) mutant may therefore offer a useful model to assist in the development of therapies for syndromes produced by altered function of human nAChRs.

  Website 
• Dale RP, Jones AK, Tamborindeguy C, Davies TG, Amey JS, Williamson S, Wolstenholme A, Field LM, Williamson MS, Walsh TK, Sattelle DB, 'Identification of ion channel genes in the Acyrthosiphon pisum genome'
  Insect Molecular Biology 19 (s2) (2010) pp.141-153
  ISSN: 0962-1075
  Abstract Aphids are major pests of crops, causing hundreds of millions of dollars worth of damage annually. Ion channel proteins are often the targets of modern insecticides and mutations in ion channel genes can lead to resistance to many leading classes of insecticides. The sequencing of the pea aphid, Acyrthosiphon pisum, genome has now allowed detailed in silico analysis of the aphid ion channels. The study has revealed significant differences in the composition of the ion channel families between the aphid and other insects. For example A. pisum does not appear to contain a homologue of the nACh receptor alpha 5 gene whilst the calcium channel beta subunit has been duplicated. These variations could result in differences in function or sensitivity to insecticides. The genome sequence will allow the study of aphid ion channels to be accelerated, leading to a better understanding of the function of these economically important channels. The potential for identifying novel insecticide targets within the aphid is now a step closer.Website 
• The International Aphid Genomics Consortium, 'Genome Sequence of the Pea Aphid Acyrthosiphon pisum'
  PLoS Biology 8 (2) (2010)
  ISSN: 1544-9173
  Abstract Aphids are important agricultural pests and also biological models for studies of insect-plant interactions, symbiosis, virus vectoring, and the developmental causes of extreme phenotypic plasticity. Here we present the 464 Mb draft genome assembly of the pea aphid Acyrthosiphon pisum. This first published whole genome sequence of a basal hemimetabolous insect provides an outgroup to the multiple published genomes of holometabolous insects. Pea aphids are host-plant specialists, they can reproduce both sexually and asexually, and they have coevolved with an obligate bacterial symbiont. Here we highlight findings from whole genome analysis that may be related to these unusual biological features. These findings include discovery of extensive gene duplication in more than 2000 gene families as well as loss of evolutionarily conserved genes. Gene family expansions relative to other published genomes include genes involved in chromatin modification, miRNA synthesis, and sugar transport. Gene losses include genes central to the IMD immune pathway, selenoprotein utilization, purine salvage, and the entire urea cycle. The pea aphid genome reveals that only a limited number of genes have been acquired from bacteria; thus the reduced gene count of Buchnera does not reflect gene transfer to the host genome. The inventory of metabolic genes in the pea aphid genome suggests that there is extensive metabolite exchange between the aphid and Buchnera, including sharing of amino acid biosynthesis between the aphid and Buchnera. The pea aphid genome provides a foundation for post-genomic studies of fundamental biological questions and applied agricultural problems.Website 
• Werren JH, Richards S, Desjardins CA, Niehuis O, Gadau J, Colbourne JK; Nasonia Genome Working Group, Werren JH, Richards S, Desjardins CA, Niehuis O, Gadau J, Colbourne JK, Beukeboom LW, Desplan C, Elsik CG, Grimmelikhuijzen CJ, Kitts P, Lynch JA, Murphy T, Oliveira DC, Smith CD, van de Zande L, Worley KC, Zdobnov EM, Aerts M, Albert S, Anaya VH, Anzola JM, Barchuk AR, Behura SK, Bera AN, Berenbaum MR, Bertossa RC, Bitondi MM, Bordenstein SR, Bork P, Bornberg-Bauer E, Brunain M, Cazzamali G, Chaboub L, Chacko J, Chavez D, Childers CP, Choi JH, Clark ME, Claudianos C, Clinton RA, Cree AG, Cristino AS, Dang PM, Darby AC, de Graaf DC, Devreese B, Dinh HH, Edwards R, Elango N, Elhaik E, Ermolaeva O, Evans JD, Foret S, Fowler GR, Gerlach D, Gibson JD, Gilbert DG, Graur D, Gründer S, Hagen DE, Han Y, Hauser F, Hultmark D, Hunter HC 4th, Hurst GD, Jhangian SN, Jiang H, Johnson RM, Jones AK, Junier T, Kadowaki T, Kamping A, Kapustin Y, Kechavarzi B, Kim J, Kim J, Kiryutin B, Koevoets T, Kovar CL, Kriventseva EV, Kucharski R, Lee H, Lee SL, Lees K, Lewis LR, Loehlin DW, Logsdon JM Jr, Lopez JA, Lozado RJ, Maglott D, Maleszka R, Mayampurath A, Mazur DJ, McClure MA, Moore AD, Morgan MB, Muller J, Munoz-Torres MC, Muzny DM, Nazareth LV, Neupert S, Nguyen NB, Nunes FM, Oakeshott JG, Okwuonu GO, Pannebakker BA, Pejaver VR, Peng Z, Pratt SC, Predel R, Pu LL, Ranson H, Raychoudhury R, Rechtsteiner A, Reese JT, Reid JG, Riddle M, Robertson HM, Romero-Severson J, Rosenberg M, Sackton TB, Sattelle DB, Schlüns H, Schmitt T, Schneider M, Schüler A, Schurko AM, Shuker DM, Simões ZL, Sinha S, Smith Z, Solovyev V, Souvorov A, Springauf A, Stafflinger E, Stage DE, Stanke M, Tanaka Y, Telschow A, Trent C, Vattathil S, Verhulst EC, Viljakainen L, Wanner KW, Waterhouse RM, Whitfield JB, Wilkes TE, Williamson M, Willis JH, Wolschin F, Wyder S, Yamada T, Yi SV, Zecher CN, Zhang L, Gibbs RA, 'Functional and evolutionary insights from the genomes of three parasitoid Nasonia species'
  Science 327 (5963) (2010) pp.343-348
  ISSN: 0036-8075 eISSN: 1095-9203
  Abstract We report here genome sequences and comparative analyses of three closely related parasitoid wasps: Nasonia vitripennis, N. giraulti, and N. longicornis. Parasitoids are important regulators of arthropod populations, including major agricultural pests and disease vectors, and Nasonia is an emerging genetic model, particularly for evolutionary and developmental genetics. Key findings include the identification of a functional DNA methylation tool kit; hymenopteran-specific genes including diverse venoms; lateral gene transfers among Pox viruses, Wolbachia, and Nasonia; and the rapid evolution of genes involved in nuclear-mitochondrial interactions that are implicated in speciation. Newly developed genome resources advance Nasonia for genetic research, accelerate mapping and cloning of quantitative trait loci, and will ultimately provide tools and knowledge for further increasing the utility of parasitoids as pest insect-control agents.Website 
• Jones A K, Bera A N, Lees K, Sattelle D B, 'The cys-loop ligand-gated ion channel gene superfamily of the parasitoid wasp, Nasonia vitripennis'
  Heredity 104 (3) (2010) pp.247-259
  ISSN: 0018-067X
  Abstract

  Members of the cys-loop ligand-gated ion channel (cysLGIC) superfamily mediate chemical neurotransmission and are studied extensively as potential targets of drugs used to treat neurological disorders, such as Alzheimer's disease. Insect cys-loop LGICs also have central roles in the nervous system and are targets of highly successful insecticides. Here, we describe the cysLGIC superfamily of the parasitoid wasp, Nasonia vitripennis, which is emerging as a highly useful model organism and is deployed as a biological control of insect pests. The wasp superfamily consists of 26 genes, which is the largest insect cysLGIC superfamily characterized, whereas Drosophila melanogaster, Apis mellifera and Tribolium castaneum have 23, 21 and 24, respectively. As with Apis, Drosophila and Tribolium, Nasonia possesses ion channels predicted to be gated by acetylcholine, γ-amino butyric acid, glutamate and histamine, as well as orthologues of the Drosophila pH-sensitive chloride channel (pHCl), CG8916 and CG12344. Similar to other insects, wasp cysLGIC diversity is broadened by alternative splicing and RNA A-to-I editing, which may also serve to generate species-specific receptor isoforms. These findings on N. vitripennis enhance our understanding of cysLGIC functional genomics and provide a useful basis for the study of their function in the wasp model, as well as for the development of improved insecticides that spare a major beneficial insect species.

  Website 
• Jones AK, Buckingham DB, Sattelle DB, 'Proteins interacting with nicotinic acetylcholine receptors: expanding functional and therapeutic horizons'
  Trends in Pharmacological Sciences 31 (10) (2010) pp.455-462
  ISSN: 0165-6147
  Abstract Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that carry out the fast actions of the neurotransmitter acetylcholine (ACh). Over the past 30 years, it has become clear that the activity of nAChRs is dependent on their interaction with a host of proteins, and the number of these that have been identified has increased considerably with recent large-scale proteomic analyses. This review focuses on these interacting proteins, discussing how they regulate a wide range of functions including receptor assembly, and trafficking to and from the cell surface, as well as how they directly modulate functional characteristics such as sensitivity and the degree of response to ACh. Mutations giving rise to disease states highlight the importance of these interacting proteins. Here, we consider their potential as future therapeutic targets for treating diseases associated with altered nAChR function.Website 
• Buckingham SD, Jones AK, Brown LA, Sattelle DB, 'Nicotinic acetylcholine receptor signalling: roles in Alzheimer’s disease and amyloid neuroprotection'
  Pharmacological Reviews 61 (1) (2009) pp.39-61
  ISSN: 0031-6997 eISSN: 1521-0081
  Abstract Alzheimer's disease (AD), the major contributor to dementia in the elderly, involves accumulation in the brain of extracellular plaques containing the β-amyloid protein (Aβ) and intracellular neurofibrillary tangles of hyperphosphorylated tau protein. AD is also characterized by a loss of neurons, particularly those expressing nicotinic acetylcholine receptors (nAChRs), thereby leading to a reduction in nAChR numbers. The Aβ_1–42 protein, which is toxic to neurons, is critical to the onset and progression of AD. The discovery of new drug therapies for AD is likely to be accelerated by an improved understanding of the mechanisms whereby Aβ causes neuronal death. We examine the evidence for a role in Aβ_1–42 toxicity of nAChRs; paradoxically, nAChRs can also protect neurons when activated by nicotinic ligands. Aβ peptides and nicotine differentially activate several intracellular signaling pathways, including the phosphatidylinositol 3-kinase/v-akt murine thymoma viral oncogene homolog pathway, the extracellular signal-regulated kinase/mitogen-activated protein kinase, and JAK-2/STAT-3 pathways. These pathways control cell death or survival and the secretion of Aβ peptides. We propose that understanding the differential activation of these pathways by nicotine and/or Aβ_1–42 may offer the prospect of new routes to therapy for AD.Website 
• Jones A K, Buckingham S D, Brown L A, Sattelle D B, 'Alternative splicing of the Anopheles Gambiae nicotine acetylcholine receptor, Agamαβ9, generates both alpha and beta subunits'
  Invertebrate Neuroscience 9 (2) (2009) pp.77-84
  ISSN: 1354-2516
  Abstract

  Nicotinic acetylcholine receptors (nAChRs) are the members of the cys-loop ligand-gated ion channel superfamily and are formed by five subunits arranged around a central ion channel. Each subunit is encoded by a separate gene and is classified as either α or non-α depending on the presence or absence, respectively, of two adjacent cysteine residues which are important for acetylcholine binding. Here, we report for the first time a single nAChR gene encoding both α and non-α subunits. Specifically, alternative splicing of the Anopheles gambiae nAChR subunit, previously called Agamα9 and renamed here Agamαβ9, generates two variants, one possessing the two cysteines (denoted Agamαβ9α) and the other lacking the cysteine doublet (Agamαβ9β). Attempts to heterologously express functional nAChRs consisting of the Agamαβ9 splice variants in Xenopus laevis oocytes were unsuccessful. Our findings further characterise a potential target to control the malaria mosquito as well as provide insights into the diversification of nAChRs.

  Website 
• Sattelle DB, Buckingham SD, Akamatsu M, Matsuda K, Pienaar I, Jones AK, Sattelle B, Almond A, Blundell CD, 'Comparative pharmacology and computational modelling yield insights into allosteric modulation of human alpha7 nicotinic acetylcholine receptors'
  Biochemical Pharmacology 78 (7) (2009) pp.836-843
  ISSN: 0006-2952
  Abstract The human α7 nicotinic acetylcholine receptor (nAChR) subunit and its Caenorhabditis elegans homolog, ACR-16, can generate functional recombinant homomeric receptors when expressed in Xenopus laevis oocytes. Both nAChRs express robustly in the presence of the co-injected chaperone, RIC-3, and show striking differences in the actions of a type I positive allosteric modulator (PAM), ivermectin (IVM). Type I PAMs are characterised by an increase in amplitude only of the response to acetylcholine (ACh), whereas type II PAMs exhibit, in addition, changes in time-course/desensitization of the ACh response. The type I PAMs, ivermectin, 5-hydroxyindole (5-HI), NS-1738 and genistein and the type II PAM, PNU-120596, are all active on human α7 but are without PAM activity on ACR-16, where they attenuate the amplitude of the ACh response. We used the published structure of avermectin B1a to generate a model of IVM, which was then docked into the candidate transmembrane allosteric binding site on α7 and ACR-16 in an attempt to gain insights into the observed differences in IVM actions. The new pharmacological findings and computational approaches being developed may inform the design of novel PAM drugs targeting major neurological disorders.Website 
• Jones A K, Buckingham S D, Papadaki M, Yokota M, Sattelle B M, Matsuda K, Sattelle D B, 'Splice-variant- and stage-specific RNA editing of the Drosophila GABA receptor modulates agonist potency'
  Journal of Neuroscience 29 (13) (2009) pp.4287-4292
  ISSN: 1529-2401 eISSN: 1529-2401
  Abstract The molecular diversity of many gene products functioning in the nervous system is enhanced by alternative splicing and adenosine-to-inosine editing of pre-mRNA. Using RDL, a Drosophila melanogaster GABA-gated ion channel, we examined the functional impact of RNA editing at several sites along with alternative splicing of more than one exon. We show that alternative splicing and RNA editing have a combined influence on the potency of the neurotransmitter GABA, and the editing isoforms detected in vivo span the entire functional range of potencies seen for all possible edit variants expressed in Xenopus laevis oocytes. The extent of RNA editing is developmentally regulated and can also be linked to the choice of alternative exons. These results provide insights into how the rich diversity of signaling necessary for complex brain function can be achieved by relatively few genes.Website 
• Buckingham SD, Kwak S, Jones AK, Blackshaw SE, Sattelle DB, 'Edited GluR2, a gatekeeper for motor neurone survival?'
  BioEssays 30 (11-12) (2008) pp.1185-1192
  ISSN: 0265-9247
  Abstract Amyotrophic lateral sclerosis (ALS) is a progressive degenerative disorder of motor neurones. Although the genetic basis of familial forms of ALS has been well explored, the molecular basis of sporadic ALS is less well understood. Recent evidence has linked sporadic ALS with the failure to edit key residues in ionotropic glutamate receptors, resulting in excessive influx of calcium ions into motor neurones which in turn triggers cell death. Here we suggest that edited AMPA glutamate (GluR2) receptor subunits serve as gatekeepers for motor neurone survivalWebsite 
• Amiri S, Shimomura M, Vijayan R, Nishiwaki H, Akamatsu M, Matsuda K, Jones AK, Sansom MS, Biggin PC, Sattelle DB, 'A role for Leu118 of loop E in agonist binding to the alpha 7 nicotinic acetylcholine receptor'
  Molecular Pharmacology 73 (6) (2008) pp.1659-1667
  ISSN: 0026-895X
  Abstract Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels mediating fast cholinergic synaptic transmission in the brain and at neuromuscular junctions. We used the structure of the acetylcholine binding protein from Lymnaea stagnalis to model the chicken α7 agonist-binding domain. The initial models and a preliminary docking study suggested that position Leu118 may play an important role in determining agonist actions on α7. A prediction from these in silico studies, that L118E and L118D would retain binding to acetylcholine but L118K and L118R would not, was confirmed in electrophysiological studies on functional recombinant mutant receptors expressed in Xenopus laevis oocytes. The functional studies also demonstrated that residues at position 118 have a dramatic effect on the actions of imidacloprid (a partial agonist of wild-type α7 receptors) and its des-nitro derivative. Molecular dynamics simulations confirmed that Leu118 can strongly influence agonist binding and that the model was robust in terms of its prediction for acetylcholine binding. Together, the results indicate a role for Leu118 in influencing agonist actions on α7 nAChRs.Website 
• Tribolium Genome Sequencing Consortium, 'The genome of the model beetle and pest Tribolium castaneum'
  Nature 452 (6784) (2008) pp.949-955
  ISSN: 0028-0836
  Abstract Tribolium castaneum is a member of the most species-rich eukaryotic order, a powerful model organism for the study of generalized insect development, and an important pest of stored agricultural products. We describe its genome sequence here. This omnivorous beetle has evolved the ability to interact with a diverse chemical environment, as shown by large expansions in odorant and gustatory receptors, as well as P450 and other detoxification enzymes. Development in Tribolium is more representative of other insects than is Drosophila, a fact reflected in gene content and function. For example, Tribolium has retained more ancestral genes involved in cell-cell communication than Drosophila, some being expressed in the growth zone crucial for axial elongation in short-germ development. Systemic RNA interference in T. castaneum functions differently from that in Caenorhabditis elegans, but nevertheless offers similar power for the elucidation of gene function and identification of targets for selective insect control.Website 
• Jones A, Sattelle D B, 'The cys-loop ligand-gated ion channel gene superfamily of the nematode, Caenorhabditis elegans'
  Invertebrate Neuroscience 8 (1) (2008) pp.41-47
  ISSN: 1354-2516
  Abstract The nematode, Caenorhabditis elegans, possesses the most extensive known superfamily of cys-loop ligand-gated ion channels (cys-loop LGICs) consisting of 102 subunit-encoding genes. Less than half of these genes have been functionally characterised which include cation-permeable channels gated by acetylcholine (ACh) and γ-aminobutyric acid (GABA) as well as anion-selective channels gated by ACh, GABA, glutamate and serotonin. Following the guidelines set for genetic nomenclature for C. elegans, we have designated unnamed subunits as lgc genes (ligand-gated ion channels of the cys-loop superfamily). Phylogenetic analysis shows that several of these lgc subunits form distinct groups which may represent novel cys-loop LGIC subtypes.Website 
• Sattelle DB, Jones AK, Buckingham SD, 'Insect genomes: challenges and opportunities for neuroscience'
  Invertebrate Neuroscience 7 (3) (2007) pp.133-136
  ISSN: 1354-2516
  Abstract EditorialWebsite 
• Jones AK, Davis P, Hodgkin J, Sattelle DB, 'The nicotinic acetylcholine receptor gene family of the nematode Caenorhabditis elegans: an update on nomenclature'
  Invertebrate Neuroscience 7 (2) (2007) pp.129-131
  ISSN: 1354-2516
  Abstract The simple nematode, Caenorhabditis elegans, possesses the most extensive known gene family of nicotinic acetylcholine receptor (nAChR)-like subunits. Whilst all show greatest similarity with nAChR subunits of both invertebrates and vertebrates, phylogenetic analysis suggests that just over half of these (32) may represent other members of the cys-loop ligand-gated ion channel superfamily. We have introduced a novel nomenclature system for these “Orphan” subunits, designating them as lgc genes (ligand-gated ion channels of the cys-loop superfamily), which can also be applied in future to unnamed and uncharacterised members of the cys-loop ligand-gated ion channel superfamily. We present here the resulting updated version of the C. elegans nAChR gene family and related ligand-gated ion channel genes.Website 
• Jones AK, Brown LA, Sattelle DB, 'Insect nicotinic acetylcholine receptor gene families: from genetic model organism to vector, pest and beneficial species'
  Invertebrate Neuroscience 7 (1) (2007) pp.67-73
  ISSN: 1354-2516
  Abstract Nicotinic acetylcholine receptors (nAChRs) mediate fast synaptic transmission in the insect nervous system and are targets of a major group of insecticides, the neonicotinoids. Analyses of genome sequences have shown that nAChR gene families remain compact in diverse insect species, when compared to their mammalian counterparts. Thus, Drosophila melanogaster and Anopheles gambiae each possess 10 nAChR genes while Apis mellifera has 11. Although these are among the smallest nAChR gene families known, receptor diversity can be considerably increased by alternative splicing and mRNA A-to-I editing, thereby generating species-specific subunit isoforms. In addition, each insect possesses at least one highly divergent nAChR subunit. Species-specific subunit diversification may offer promising targets for future rational design of insecticides that act on particular pests while sparing beneficial insects. Electrophysiological studies on cultured Drosophila cholinergic neurons show partial agonist actions of the neonicotinoid imidacloprid and super-agonist actions of another neonicotinoid, clothianidin, on native nAChRs. Recombinant hybrid heteromeric nAChRs comprising Drosophila Dα2 and a vertebrate β2 subunit have been instructive in mimicking such actions of imidacloprid and clothianidin. Unitary conductance measurements on native nAChRs indicate that more frequent openings of the largest conductance state may offer an explanation for the superagonist actions of clothianidin.Website 
• Bentley GN, Jones AK, Agnew A, 'ShAR2beta, a divergent nicotinic acetylcholine receptor subunit from the blood fluke Schistosoma'
  Parasitology 134 (6) (2007) pp.833-840
  ISSN: 0031-1820 eISSN: 1469-8161
  Abstract Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that mediate the fast actions of the neurotransmitter, acetylcholine. Invertebrate nAChRs are of interest as they are targets of widely-selling insecticides and drugs that control nematode parasites. Here, we report the cloning of ShAR2β, a candidate nAChR subunit from the blood fluke, Schistosoma haematobium, which is the third trematode nAChR subunit to be characterized. While ShAR2β possesses key structural features common to all nAChRs, its amino acid sequence shares considerably low identity with those of insect, nematode and vertebrate nAChR subunits. In particular, the second transmembrane domain of ShAR2β, which lines the ion channel, bears unusual amino acid residues which will likely give rise to a receptor with distinct functional properties. Phylogenetic analysis shows that ShAR2β is a divergent nAChR subunit that may define a clade of trematode-specific subunits. We discuss our findings in the context of potentially exploiting this receptor as a target for controlling schistosome parasites.Website 
• Jones AK, Raymond-Delpech V, Thany SH, Gauthier M, Sattelle DB, 'The nicotinic acetylcholine receptor gene family of the honey bee, Apis mellifera'
  Genome Research 16 (2006) pp.1422-1430
  ISSN: 1088-9051 eISSN: 1549-5469
  Abstract Nicotinic acetylcholine receptors (nAChRs) mediate fast cholinergic synaptic transmission and play roles in many cognitive processes. They are under intense research as potential targets of drugs used to treat neurodegenerative diseases and neurological disorders such as Alzheimer's disease and schizophrenia. Invertebrate nAChRs are targets of anthelmintics as well as a major group of insecticides, the neonicotinoids. The honey bee, Apis mellifera, is one of the most beneficial insects worldwide, playing an important role in crop pollination, and is also a valuable model system for studies on social interaction, sensory processing, learning, and memory. We have used the A. mellifera genome information to characterize the complete honey bee nAChR gene family. Comparison with the fruit fly Drosophila melanogaster and the malaria mosquito Anopheles gambiae shows that the honey bee possesses the largest family of insect nAChR subunits to date (11 members). As with Drosophila and Anopheles, alternative splicing of conserved exons increases receptor diversity. Also, we show that in one honey bee nAChR subunit, six adenosine residues are targeted for RNA A-to-I editing, two of which are evolutionarily conserved in Drosophila melanogaster and Heliothis virescens orthologs, and that the extent of editing increases as the honey bee lifecycle progresses, serving to maximize receptor diversity at the adult stage. These findings on Apis mellifera enhance our understanding of nAChR functional genomics and provide a useful basis for the development of improved insecticides that spare a major beneficial insect species.Website 
• Honeybee Genome Sequencing Consortium, 'Insights into social insects from the genome of the honeybee Apis mellifera'
  Nature (2006)
  ISSN: 0028-0836 eISSN: 1476-4687
  Website 
• Jones AK, Sattelle DB, 'The cys-loop ligand-gated ion channel superfamily of the honeybee, Apis mellifera'
  Invertebrate Neuroscience 6 (3) (2006) pp.123-132
  ISSN: 1354-2516
  Abstract Members of the cys-loop ligand-gated ion channel (cys-loop LGIC) superfamily mediate neurotransmission in insects and are targets of successful insecticides. We have described the cys-loop LGIC superfamily of the honeybee, Apis mellifera, which is an important crop pollinator and a key model for social interaction. The honeybee superfamily consists of 21 genes, which is slightly smaller than that of Drosophila melanogaster comprising 23 genes. As with Drosophila, the honeybee possesses ion channels gated by acetylcholine, γ-amino butyric acid, glutamate and histamine as well as orthologs of the Drosophila pH-sensitive chloride channel (pHCl), CG8916, CG12344 and CG6927. Similar to Drosophila, honeybee cys-loop LGIC diversity is broadened by differential splicing which may also serve to generate species-specific receptor isoforms. These findings on Apis mellifera enhance our understanding of cys-loop LGIC functional genomics and provide a useful basis for the development of improved insecticides that spare a major beneficial insect species.Website 
• Brown LA, Jones AK, Buckingham SD, Mee CJ, Sattelle DB, 'Contributions from Caenorhabditis elegans functional genetics to antiparasitic drug target identification and validation: Nicotinic acetylcholine receptors, a case study'
  International Journal for Parasitology 36 (6) (2006) pp.617-624
  ISSN: 0020-7519
  Abstract Following the complete sequencing of the genome of the free-living nematode, Caenorhabditis elegans, in 1998, rapid advances have been made in assigning functions to many genes. Forward and reverse genetics have been used to identify novel components of synaptic transmission as well as determine the key components of antiparasitic drug targets. The nicotinic acetylcholine receptors (nAChRs) are prototypical ligand-gated ion channels. The functions of these transmembrane proteins and the roles of the different members of their extensive subunit families are increasingly well characterised. The simple nervous system of C. elegans possesses one of the largest nicotinic acetylcholine receptor gene families known for any organism and a combination of genetic, microarray, physiological and reporter gene expression studies have added greatly to our understanding of the components of nematode muscle and neuronal nAChR subtypes. Chemistry-to-gene screens have identified five subunits that are components of nAChRs sensitive to the antiparasitic drug, levamisole. A novel, validated target acting downstream of the levamisole-sensitive nAChR has also been identified in such screens. Physiology and molecular biology studies on nAChRs of parasitic nematodes have also identified levamisole-sensitive and insensitive subtypes and further subdivisions are under investigation.Website 
• Jones AK, Marshall J, Blake AD, Buckingham SD, Darlison MG, Sattelle DB, 'Sgbeta1, a novel locust (Schistocerca gregaria) non-alpha nicotinic acetylcholine receptor-like subunit with homology to the Drosophila melanogaster Dbeta1 subunit'
  Invertebrate Neuroscience 5 (3) (2005) pp.147-155
  ISSN: 1354-2516
  Abstract The cloning, sequencing and functional expression of Sgβ1, a novel locust (Schistocerca gregaria) non-α nicotinic acetylcholine receptor (nAChR) subunit is described. This subunit shows 80% identity with the Drosophila melanogaster Dβ1 and 92% identity with the Locusta migratoria β1, non-α subunits but only 38% identity to Sgα1 (also referred to as αL1), a previously cloned S. gregaria nAChR α-subunit. When expressed in Xenopus laevis oocytes, Sgβ1 does not respond to nicotine. Responses to nicotine are observed, however, in oocytes co-expressing Sgα1 and Sgβ1, but the pharmacology is indistinguishable from that of currents produced by expressing Sgα1 alone. We conclude that either Sgβ1 does not co-assemble with Sgα1, or that it is unable to contribute to the functional properties of the receptor, in the Xenopus oocyte expression system.Website 
• Bentley GN, Jones AK, Agnew A, 'Expression and comparative functional characterisation of recombinant acetylcholinesterase from three species of Schistosoma'
  Molecular and Biochemical Parasitology 141 (1) (2005) pp.119-123
  ISSN: 0166-6851 eISSN: 1872-9428
  Abstract Short communicationWebsite 
• Jones AK, Buckingham SD, Sattelle DB, 'Chemistry-to-gene screens in Caenorhabditis elegans'
  Nature Reviews Drug Discovery 4 (2005) pp.321-330
  ISSN: 1474-1776 eISSN: 1474-1784
  Abstract The nematode worm Caenorhabditis elegans is a genetic model organism linked to an impressive portfolio of fundamental discoveries in biology. This free-living nematode, which can be easily and inexpensively grown in the laboratory, is also a natural vehicle for screening for drugs that are active against nematode parasites. Here, we show that chemistry-to-gene screens using this animal model can define targets of antiparasitic drugs, identify novel candidate drug targets and contribute to the discovery of new drugs for treating human diseases.Website 
• Sattelle DB, Jones AK, Sattelle BM, Matsuda K, Reenan R, Biggin PC, 'Edit, cut and paste in the nicotinic acetylcholine receptor gene family of Drosophila melanogaster'
  BioEssays 27 (4) (2005) pp.366-376
  ISSN: 0265-9247
  Abstract Nicotinic acetylcholine receptors (nAChRs) are important for fast synaptic cholinergic transmission. They are targets of drugs/chemicals for human and animal health as well as for pest control. With the advent of genome sequencing, entire nAChR gene families have now been described for vertebrates and invertebrates. Mostly, these are extensive with a large number of distinct subunits, making possible many nAChR subtypes differing in transmitter affinity, channel conductance, ion selectivity, desensitization, modulation and pharmacology. The smallest nAChR gene family to date is that of the fruit fly, Drosophila melanogaster, with only 10 members. This apparently compact family belies its true diversity as 4 of the 10 subunits show alternative splicing. Also, using Drosophila, A-to-I pre-mRNA editing has been demonstrated for the first time in nAChRs. Such is the extent of this variation, that one subunit alone (Dα6) can potentially generate far more isoforms than seen in entire gene families from other species. We present here three-dimensional models constructed for insect nAChRs, which show that many variations introduced by alternative splicing and RNA editing may influence receptor functionWebsite 
• Jones AK, Grauso M, Sattelle DB, 'The nicotinic acetylcholine receptor gene family of the malaria mosquito, Anopheles gambiae'
  Genomics 85 (2) (2005) pp.176-187
  ISSN: 0888-7543 eISSN: 1089-8646
  Abstract Nicotinic acetylcholine receptors (nAChRs) mediate fast cholinergic synaptic transmission in the insect nervous system and are targets of widely selling insecticides. We have identified the nAChR gene family from the genome of the malaria mosquito vector, Anopheles gambiae, to be the second complete insect nAChR gene family described following that of Drosophila melanogaster. Like Drosophila, Anopheles possesses 10 nAChR subunits with orthologous relationships evident between the two insects. Interestingly, the Anopheles orthologues of Dβ2 and Dβ3 possess the vicinal cysteines that define α subunits. As with Dα4 and Dα6, the Anopheles orthologues are alternatively spliced at equivalent exons. Reverse transcription-polymerase chain reaction analysis shows that RNA A-to-I editing sites conserved between Dα6 of Drosophila and α7-2 of the tobacco budworm, Heliothis virescens, are not shared with the equivalent nAChR subunit of Anopheles. Indeed, RNA-editing sites identified in functionally significant regions of Dβ1, Dα5, and Dα6 are not conserved in the mosquito orthologues, indicating considerable divergence of RNA molecules targeted for editing within the insect order Diptera. These findings shed further light on the diversity of nAChR subunits and may present a useful basis for the development of improved malaria control agents by enhancing our understanding of a validated mosquito insecticide target.Website 
• Buckingham SD, Pym L, Jones AK, Brown L, Sansom MS, Sattelle DB, Biggin PC, 'A7DB: A relational database for mutational, physiological and pharmacological data related to the alpha7 nicotinic acetylcholine receptor'
  BMC Neuroscience 6 (2) (2005)
  ISSN: 1471-2202
  Abstract Background
  Nicotinic acetylcholine receptors (nAChRs) are pentameric proteins that are important drug targets for a variety of diseases including Alzheimer's, schizophrenia and various forms of epilepsy. One of the most intensively studied nAChR subunits in recent years has been α7. This subunit can form functional homomeric pentamers (α7)5, which can make interpretation of physiological and structural data much simpler. The growing amount of structural, pharmacological and physiological data for these receptors indicates the need for a dedicated and accurate database to provide a means to access this information in a coherent manner.
   
  Description
  A7DB http://www.lgics.org/a7db/ is a new relational database of manually curated experimental physiological data associated with the α7 nAChR. It aims to store as much of the pharmacology, physiology and structural data pertaining to the α7 nAChR. The data is accessed via web interface that allows a user to search the data in multiple ways: 1) a simple text query 2) an incremental query builder 3) an interactive query builder and 4) a file-based uploadable query. It currently holds more than 460 separately reported experiments on over 85 mutations.
   
  Conclusions
  A7DB will be a useful tool to molecular biologists and bioinformaticians not only working on the α7 receptor family of proteins but also in the more general context of nicotinic receptor modelling. Furthermore it sets a precedent for expansion with the inclusion of all nicotinic receptor families and eventually all cys-loop receptor families.
  Website 
• Culetto E, Baylis HA, Richmond JE, Jones AK, Fleming JT, Squire MD, Lewis JA, Sattelle DB, 'The Caenorhabditis elegans unc-63 gene encodes a levamisole-sensitive nicotinic acetylcholine receptor alpha subunit'
  Journal of Biological Chemistry 279 (2004) pp.42476-42483
  ISSN: 0021-9258
  Abstract The anthelmintic drug levamisole causes hypercontraction of body wall muscles and lethality in nematode worms. In the nematode Caenorhabditis elegans, a genetic screen for levamisole resistance has identified 12 genes, three of which (unc-38, unc-29, and lev-1) encode nicotinic acetylcholine receptor (nAChR) subunits. Here we describe the molecular and functional characterization of another levamisole-resistant gene, unc-63, encoding a nAChR α subunit with a predicted amino acid sequence most similar to that of UNC-38. Like UNC-38 and UNC-29, UNC-63 is expressed in body wall muscles. In addition, UNC-63 is expressed in vulval muscles and neurons. We also show that LEV-1 is expressed in body wall muscle, thus overlapping the cellular localization of UNC-63, UNC-38, and UNC-29 and suggesting possible association in vivo. This is supported by electrophysiological studies on body wall muscle, which demonstrate that a levamisole-sensitive nAChR present at the C. elegans neuromuscular junction requires both UNC-63 and LEV-1 subunits. Thus, at least four subunits, two α types (UNC-38 and UNC-63) and two non-α types (UNC-29 and LEV-1), can contribute to levamisole-sensitive muscle nAChRs in nematodes.Website 
• Bentley GN, Jones AK, Oliveros Parra WG, Agnew A, 'ShAR1alpha and ShAR1beta: novel putative nicotinic acetylcholine receptor subunits from the platyhelminth blood fluke Schistosoma'
  Gene 329 (2004) pp.27-38
  ISSN: 0378-1119 eISSN: 1879-0038
  Abstract The cDNAs for two novel neuronal-type nicotinic acetylcholine receptor (nAChR) subunits have been cloned and characterised from the parasitic trematode blood fluke Schistosoma haematobium. One of these encodes a putative nAChR α-subunit named ShAR1α, whilst the second encodes a potential non-α subunit, ShAR1β. These ShARs possess the key structural features common to all nAChRs, but they are unusual in that they have very large cytoplasmic domains spanning M3 and M4. Overall, the ShAR1α and ShAR1β proteins share 37% identity and 53% similarity, but excluding the residues of the M3–M4 domain this rises to 52% identity and 71% similarity. Sequence comparisons with other nAChR polypeptides indicate that both ShARs are most similar to the invertebrate α7-like subunits identified in insects and nematodes, and to the vertebrate subunits α7 and α8. Outside of the M3–M4 domain, 45% and 40%, respectively, of the ShAR1α and ShAR1β residues are conserved in the ACR-16 subunit from Caenorhabditis elegans. Phylogenetic analysis suggests that the ShARs share a common lineage with members of the ACR-16 group as well as α7 and α8. Immunolocalisation studies revealed distinct and non-overlapping patterns of distribution for ShAR1α and ShAR1β within the parasite. ShAR1β was localised within the musculature and on discrete cell bodies within the connective parenchyma. In contrast, ShAR1α was localised exclusively to the surface membranes, suggesting it may contribute to the regulatory nAChR we have characterised previously. In Xenopus oocyte expression studies, ShAR1α did not form functional channels on its own or in combination with ShAR1β or the chick β2 subunit. Furthermore, a chimera in which the M3–M4 domain of ShAR1α was replaced with that of chick α7 was also non-functional. We discuss our findings in the context of the proposed role for surface nAChRs in the regulation of glucose uptake in the parasite, and the potential exploitation of these receptors as targets for cholinergic schistosomicides.Website 
• Jones AK, Sattelle DB, 'Functional genomics of the nicotinic acetylcholine receptor family of the nematode Caenorhabditis elegans'
  BioEssays 26 (1) (2003) pp.39-49
  ISSN: 0265-9247
  Abstract Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that bring about a diversity of fast synaptic actions. Analysis of the Caenorhabditis elegans genome has revealed one of the most-extensive and diverse nAChR gene families known, consisting of at least 27 subunits. Striking variation with possible functional implications has been observed in normally conserved motifs at the acetylcholine-binding site and in the channel-lining region. Some nAChR subunits are particular to neurons whilst others are present in both neurons and muscles. The localization of subunits in non-synaptic regions suggests novel roles for nAChRs. Genetic and heterologous expression studies have identified a subset of nAChR subunits that are important drug targets while the study of mutants has identified genes functionally-linked to nAChRs. Future studies using C. elegans offer the prospect of increasing our understanding of the functional diversity of a complex nAChR gene family as well as addressing the role of nAChRs and associated proteins in human disorders.Website 
• Jones AK, Elgar G, Sattelle DB, 'The nicotinic acetylcholine receptor gene family of the pufferfish, Fugu rubripes'
  Genomics 82 (4) (2003) pp.441-451
  ISSN: 0888-7543 eISSN: 1089-8646
  Abstract Nicotinic acetylcholine receptors (nAChRs) mediate fast cholinergic synaptic transmission at nerve–muscle junctions and in the brain. However, the complete gene family of nAChRs has not so far been reported for any vertebrate organism. We have identified the complete nAChR gene family from the reference genome of the pufferfish, Fugu rubripes. It consists of 16 α and 12 non-α candidate subunits, making it the largest vertebrate nAChR gene family known to date. The gene family includes an unusual set of muscle-like nAChR subunits comprising two α1s, two β1s, one δ, one ε, and one γ. One of the β1 subunits possesses an aspartate residue and N-glycosylation sites hitherto shown to be necessary for δ-subunit function. Potential Fugu orthologs of neuronal nAChR subunits α2–4, α6, and β2–4 have been identified. Interestingly, the Fugu α5 counterpart appears to be a non-α subunit. Fugu possesses an expanded set of α7–9-like subunits and no α10 ortholog has been found. Two new candidate β subtypes, designated β5 and β6, may represent subunits yet to be found in the human genome. The Fugu nAChR gene structures are considerably more diverse than those of higher vertebrates, with evidence of “intron gain” in many cases. We show, using RT-PCR, that the Fugu nAChR subunits are expressed in a variety of tissues.Website 
• Bentley GN, Jones AK, Agnew A, 'Mapping and sequencing of acetylcholinesterase genes from the platyhelminth blood fluke Schistosoma'
  Gene 314 (2003) pp.103-112
  ISSN: 0378-1119 eISSN: 1879-0038
  Abstract Acetylcholinesterase (AChE) on the surface of the parasitic blood fluke Schistosoma is the likely target for schistosomicidal anticholinesterases. Determination of the molecular structure of this drug target is key for the development of improved anticholinesterase drugs and potentially a novel vaccine. We have recently cloned the cDNA encoding the AChE from the human parasite Schistosoma haematobium and succeeded in expressing functional recombinant protein. We now describe the cloning and molecular characterisation of homologues from two other schistosome species—Schistosoma mansoni and Schistosoma bovis, which are important parasites of man and cattle, respectively, but which differ in their sensitivity to the therapeutic anticholinesterase metrifonate. Comparison of the deduced amino acid sequences revealed that the AChE from all three species posses a high degree of identity, with conservation of all of the residues known to be important for substrate binding and catalytic activity. Also conserved is a unique C-terminal domain which is unusual in that it lacks the consensus for GPI modification, even though the native protein is considered to be GPI-anchored. We have also established the AChE gene structures for all three species and cloned the complete gene for S. haematobium AChE. The gene structure is relatively complex, comprising nine coding exons; the location of the splice sites is identical in all three species, but the size of the introns varies considerably. The two C-terminal splicing sites that are conserved in all species are also present in Schistosoma, but a third C-terminal conserved splicing site which is located 11–13 amino acids upstream of the histidine of the catalytic triad in all invertebrate AChE genes characterised to date is absent. We discuss our findings in the context of the molecular phylogeny of the AChE genes and the potential application to the control of schistosomiasis.Website 
• Mongan NP, Jones AK, Smith GR, Sansom MS, Sattelle DB, 'Novel alpha7-like nicotinic acetylcholine receptor subunits in the nematode Caenorhabditis elegans'
  Protein Science 11 (5) (2002) pp.1162-1171
  ISSN: 0961-8368
  Abstract We have used reverse-transcription-polymerase chain reaction (RT-PCR) and DNA sequencing techniques to confirm the transcription of seven (six α and one non-α) novel candidate nicotinic acetylcholine receptor (nAChR) subunit-encoding genes identified in the genome sequence of the nematode Caenorhabditis elegans. Compared to vertebrate nAChR subunits, they most closely resemble the homomer-forming, neuronal α7 subunit. Comparison of the predicted amino acid sequences of the new nAChR subunits with those described previously in C. elegans reveals five subunits (four α and one non-α) which resemble the DEG-3-like group of subunits. To date, this highly divergent nAChR subunit group is unique to C. elegans. ACR-22 is the first non-α member of the DEG-3-like group of subunits to be identified. Two new members of the related ACR-16-like nAChR group of subunits have also been shown to be transcribed, making the ACR-16-like subunit group the largest in C. elegans. Residues in the α subunit second transmembrane region (M2) which contribute to the channel lining show variations with implications for channel function. For example, in ACR-22, the highly conserved 0′ lysine of M2 is replaced by histidine. Restrained molecular dynamics simulations have been used to generate molecular models of homo-pentameric M2 helix bundles for the novel subunits, enabling identification and display of pore-lining and protein interface residues. The number and diversity of genes encoding C. elegans nAChR subunits with similarities to the homomer-forming vertebrate α7 subunits and the identification of related non-α subunits, only found in C. elegans to date, suggest that at least some of these subunits may contribute to heteromers in vivo.Website 
• Jones AK, Bentley GN, Oliveros Parra WG, Agnew A, 'Molecular characterisation of an acetylcholinesterase implicated in the regulation of glucose scavenging by the parasite Schistosoma'
  FASEB Journal (2002)
  ISSN: 0892-6638 eISSN: 1530-6860
  Website 

Book chapters

• Taylor-Wells J, Jones AK, 'Variations in the insect GABA receptor, RDL, and their impact on receptor pharmacology' in Gross AD, Ozoe Y, Coats JR (ed.), Advances in agrochemicals: ion channels and G Protein-Coupled receptors (GPCRs) as Targets for Pest Control. Volume 2, GPCRs and Ion Channels. ACS Symposium Series, ACS Publications (2017)
  ISBN: 9780841232600 eISBN: 9780841232587
  Abstract

  The resistance to dieldrin (RDL) receptor is an insect γ-aminobutyric acid (GABA) receptor, characterized by the dieldrin resistance mutation that was pivotal to understanding target based insecticide resistance. RDL is the target for various non-competitive antagonists, including dieldrin and fipronil, as well as novel acting compounds such as the meta-diamides and isoxazolines. Therefore the RDL receptor has returned to center stage as a relevant and effective insecticide target. Our understanding of the function of RDL in vivo is still unfolding, with the discovery of species specific post-transcriptional modifications such as alternative splicing and RNA editing, modifications shown to influence the pharmacology of the receptor. Exposing these receptors to insecticides also evokes ever evolving mechanisms of mutagenesis, and a number of contributory mutations have been identified both in field and laboratory resistant insects, occurring in parallel to the dieldrin resistance mutation. We present an overview of these variations and discuss the impact on the pharmacology of GABA and various insecticides.
  

  Website 
• Jones AK, Sattelle DB, 'Diversity of Insect Nicotinic Acetylcholine Receptor Subunits' in Thany S (ed.), Insect Nicotinic Acetylcholine Receptors, Landes Bioscience (2010)
  ISBN: 978-1-4419-6445-8
  Abstract Nicotinic acetylcholine receptors (nAChRs) are ligand-gated ion channels that mediate fast synaptic transmission in the insect nervous system and are targets of a major group of insecticides, the neonicotinoids. They consist of five subunits arranged around a central ion channel. Since the subunit composition determines the functional and pharmacological properties of the receptor the presence of nAChR families comprising several subunit-encoding genes provides a molecular basis for broad functional diversity. Analyses of genome sequences have shown that nAChR gene families remain compact in diverse insect species, when compared to their nematode and vertebrate counterparts. Thus, the fruit fly (Drosophila melanogaster), malaria mosquito (Anopheles gambiae), honey bee (Apis mellifera), silk worm (Bombyx mori) and the red flour beetle (Tribolium castaneum) possess 10–12 nAChR genes while human and the nematode Caenorhabditis elegans have 16 and 29 respectively. Although insect nAChR gene families are amongst the smallest known, receptor diversity can be considerably increased by the posttranscriptional processes alternative splicing and mRNA A-to-I editing which can potentially generate protein products which far outnumber the nAChR genes. These two processes can also generate species-specific subunit isoforms. In addition, each insect possesses at least one highly divergent nAChR subunit which may perform species-specific functions. Species-specific subunit diversification may offer promising targets for future rational design of insecticides that target specific pest insects while sparing beneficial species.Website 
• Sattelle DB, Jones AK, Brown LA, Buckingham SD, Mee CJ, Pym L, 'Nicotinic acetylcholine receptors as drug/chemical targets, contributions from comparative genomics, forward and reverse genetics' in Parrington J, Coward K (ed.), Comparative Genomics and Proteomics in Drug Discovery, Taylor and Francis Group (2006)
  ISBN: 9780415396530
  Abstract Chapter 4
• Sattelle DB, Culetto E, Jones AK, 'Molecular and functional diversity in the nicotinic acetylcholine receptor gene families of Caenorhabditis elegans and Drosophila melanogaster' in Fisher A, Soreq H (ed.), Cholinergic Mechanism – Function and Dysfunction, Taylor and Francis (2004)
  ISBN: 9781841840758
  Abstract Chapter 31Website 

Quotations in the press:

Independent: Bee-friendly insecticides closer to reality after breakthrough development

Press, publicity and reviews

Undergraduate project students in the laboratory of Andrew Jones are analysing mosquito DNA to determine the extent of insecticide resistance in Laos PDR, where Dengue fever and malaria are important health concerns:

https://www.brookes.ac.uk/about-brookes/news/mosquito-dna-study-offers-crucial-clues-to-disease-control-in-laos/

Further information

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Twitter:Andrew Jones@AKJBrookes

ResearchGate: https://www.researchgate.net/profile/Andrew_Jones26

Google Scholar:https://scholar.google.co.uk/citations?hl=en&user=rWhzuN8AAAAJ