Glutamate-Gated Chloride Channels of Haemonchus contortus Restore Drug Sensitivity to Ivermectin Resistant Caenorhabditis elegans
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{"title"=>"Glutamate-Gated chloride channels of Haemonchus contortus restore drug sensitivity to ivermectin resistant Caenorhabditis elegans", "type"=>"journal", "authors"=>[{"first_name"=>"Susan K.", "last_name"=>"Glendinning", "scopus_author_id"=>"48662132600"}, {"first_name"=>"Steven D.", "last_name"=>"Buckingham", "scopus_author_id"=>"7006313456"}, {"first_name"=>"David B.", "last_name"=>"Sattelle", "scopus_author_id"=>"7006226532"}, {"first_name"=>"Susan", "last_name"=>"Wonnacott", "scopus_author_id"=>"7006057996"}, {"first_name"=>"Adrian J.", "last_name"=>"Wolstenholme", "scopus_author_id"=>"7003753272"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"79960777044", "pmid"=>"21818319", "isbn"=>"1932-6203", "scopus"=>"2-s2.0-79960777044", "issn"=>"19326203", "pui"=>"362223682", "doi"=>"10.1371/journal.pone.0022390"}, "id"=>"344ec333-9d98-3c63-9f6f-50579bcad6e3", "abstract"=>"Anthelmintic resistance is a major problem in livestock farming, especially of small ruminants, but our understanding of it has been limited by the difficulty in carrying out functional genetic studies on parasitic nematodes. An important nematode infecting sheep and goats is Haemonchus contortus; in many parts of the world this species is resistant to almost all the currently available drugs, including ivermectin. It is extremely polymorphic and to date it has proved impossible to relate any sequence polymorphisms to its ivermectin resistance status. Expression of candidate drug-resistance genes in Caenorhabditis elegans could provide a convenient means to study the effects of polymorphisms found in resistant parasites, but may be complicated by differences between the gene families of target and model organisms. We tested this using the glutamate-gated chloride channel (GluCl) gene family, which forms the ivermectin drug target and are candidate resistance genes. We expressed GluCl subunits from C. elegans and H. contortus in a highly resistant triple mutant C. elegans strain (DA1316) under the control of the avr-14 promoter; expression of GFP behind this promoter recapitulated the pattern previously reported for avr-14. Expression of ivermectin-sensitive subunits from both species restored drug sensitivity to transgenic worms, though some quantitative differences were noted between lines. Expression of an ivermectin-insensitive subunit, Hco-GLC-2, had no effect on drug sensitivity. Expression of a previously uncharacterised parasite-specific subunit, Hco-GLC-6, caused the transgenic worms to become ivermectin sensitive, suggesting that this subunit also encodes a GluCl that responds to the drug. These results demonstrate that both orthologous and paralogous subunits from C. elegans and H. contortus are able to rescue the ivermectin sensitivity of mutant C. elegans, though some quantitative differences were observed between transgenic lines in some assays. C. elegans is a suitable system for studying parasitic nematode genes that may be involved in drug resistance.", "link"=>"http://www.mendeley.com/research/glutamategated-chloride-channels-haemonchus-contortus-restore-drug-sensitivity-ivermectin-resistant", "reader_count"=>51, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Researcher"=>13, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>17, "Other"=>4, "Student > Master"=>9, "Student > Bachelor"=>2, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>1, "Researcher"=>13, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>17, "Other"=>4, "Student > Master"=>9, "Student > Bachelor"=>2, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>5, "Engineering"=>1, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>6, "Agricultural and Biological Sciences"=>33, "Medicine and Dentistry"=>1, "Veterinary Science and Veterinary Medicine"=>3, "Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>33}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}, "Unspecified"=>{"Unspecified"=>5}, "Environmental Science"=>{"Environmental Science"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>3}}, "reader_count_by_country"=>{"Canada"=>1, "Colombia"=>1, "United States"=>2, "Brazil"=>1, "United Kingdom"=>1, "Mexico"=>1, "Kenya"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/752278"], "description"=>"<p>The <i>avr-14</i> promoter fragment used was a section 1.7 kb upstream of the initiation codon. A) Whole adult wild type <i>C. elegans</i> transformed with a <i>Cel-avr-14::gfp</i> construct. vnc  =  ventral nerve cord; nr  =  nerve ring. B) Head section. an  =  amphidial neurones; vnc  =  ventral nerve cord. C) Body section between vulva and tail. pds  =  posterior deirid sensilla neurone cell bodies; vnc  =  ventral nerve cord. D) Tail section. phn  =  PHA, PHB and PHC neurone cell bodies. vnc  =  ventral nerve cord.</p>", "links"=>[], "tags"=>["gfp"], "article_id"=>422645, "categories"=>["Genetics", "Infectious Diseases"], "users"=>["Susan K. Glendinning", "Steven D. Buckingham", "David B. Sattelle", "Susan Wonnacott", "Adrian J. Wolstenholme"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022390.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_pattern_of_GFP_under_the_control_of_the_avr_14_promoter_/422645", "title"=>"Expression pattern of GFP under the control of the <i>avr-14</i> promoter.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:21:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/752415"], "description"=>"<p>N = 3. Asterisks indicate a significant difference (p = <0.05) between the transgenic lines and the parent DA1316 triple mutant (<i>avr-14; avr-15; glc-1</i>). A) Ivermectin sensitivity, assessed by counting the number of stationary worms after exposure to 1 µM ivermectin for 1 hr, of DA1316 transformed with either <i>Cel-avr-14a</i> (3 lines) or <i>Cel-avr-14b</i> (2 lines) cDNA. B) Ivermectin sensitivity of DA1316 transformed with <i>Hco-avr-14b</i> cDNA (4 lines), assessed as in panel A). For panels A & B, data are presented as mean ± SEM and a one-way ANOVA and Tukey's post-hoc test were performed on the two data sets separately using Minitab. Wild-type N2 worms and the DA1370 double mutant (<i>avr-15; glc-1</i>) are included for comparison. C) The effect of ivermectin on nematode swimming measured as “thrashes” per minute (as % of vehicle control) for wild type and transgenic <i>C. elegans</i> lines. <i>Hco-avr-14b</i> in DA1316 line 2 and <i>Cel-avr-14b</i> in DA1316 line 1 were chosen as representative lines from the motility assay for use in the thrashing assays. The experiment was repeated 3 times, with 8 worms per strain per concentration. D) An automated thrashing assay on the same lines as in panel C. The experiment was repeated between 1 and 4 times for each data point. Curves were fitted using the variable slope sigmoidal equation in GraphPad Prism (San Diego). The data are presented as mean ± SEM with N = 3. Symbols as in panel C).</p>", "links"=>[], "tags"=>["assays", "ivermectin", "cdnas", "resistant", "da1316"], "article_id"=>422786, "categories"=>["Genetics", "Infectious Diseases"], "users"=>["Susan K. Glendinning", "Steven D. Buckingham", "David B. Sattelle", "Susan Wonnacott", "Adrian J. Wolstenholme"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022390.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Motility_assays_showing_rescue_of_ivermectin_sensitivity_by_avr_14_cDNAs_in_the_resistant_DA1316_mutant_/422786", "title"=>"Motility assays showing rescue of ivermectin sensitivity by <i>avr-14</i> cDNAs in the resistant DA1316 mutant.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:22:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/752496"], "description"=>"<p>A). The number of eggs that developed to adulthood on different concentrations of ivermectin. There is rescue of ivermectin sensitivity by <i>Cel-avr-14a</i>, <i>Cel-avr-14b</i> and <i>Hco-avr-14b</i> cDNA in the resistant DA1316 mutant. Data are presented as mean ± SEM. B) The DA1316 triple mutant transformed with <i>Hco-avr-14b</i> cDNA carried out fewer reversals per minute than the other lines. Reversals per minute were counted to compare WT, DA1316 (<i>glc-1</i>; <i>avr-14</i>; <i>avr-15</i>) and DA1316 containing either <i>Cel-avr-14b</i> or <i>Hco-avr-14b</i> cDNA behind the <i>Cel-avr-14</i> promoter. The double mutant, DA1370 (<i>avr-15; glc-1</i>), which has mutations in <i>glc-1</i> and <i>avr-15</i> was also used as a comparison. * Indicates that the reversals of the line containing the cDNA from the parasite were significantly reduced compared to DA1316, WT and DA1316 containing <i>Cel-avr-14b</i> cDNA. No difference was detected between the reversal frequencies of WT, DA1316, DA1370 or <i>Cel-avr-14b</i> in DA1316, using a one-way ANOVA (Minitab). Data are presented as mean ± SEM with N = 10 worms for each line.</p>", "links"=>[], "tags"=>["behavioural", "assays", "demonstrating"], "article_id"=>422866, "categories"=>["Genetics", "Infectious Diseases"], "users"=>["Susan K. Glendinning", "Steven D. Buckingham", "David B. Sattelle", "Susan Wonnacott", "Adrian J. Wolstenholme"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022390.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Development_and_behavioural_assays_demonstrating_rescue_of_the_avr_14_phenotypes_/422866", "title"=>"Development and behavioural assays demonstrating rescue of the <i>avr-14</i> phenotypes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:22:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/752563"], "description"=>"<p>A) Paralysis assay on lines transformed with cDNA expressing other <i>C. elegans</i> GluCl subunits. The proportion of worms that were paralysed by immersion in 1 µM ivermectin for 1 hr is shown. B) Thrashing assay of the lines from panel A. The number of thrashes/minute of the worms following a 1 hr exposure to 1 µM ivermectin is indicated. C) Paralysis assay on worms transformed with cDNAs expressing <i>H. contortus</i> GluCl subunits. D) Thrashing assay at three different concentrations of ivermectin, carried out on some of the lines shown in C). For panels A, B & C, *  =  statistically significant difference from DA1316 (p≤0.05).</p>", "links"=>[], "tags"=>["ivermectin-sensitive", "subunits"], "article_id"=>422933, "categories"=>["Genetics", "Infectious Diseases"], "users"=>["Susan K. Glendinning", "Steven D. Buckingham", "David B. Sattelle", "Susan Wonnacott", "Adrian J. Wolstenholme"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022390.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Other_ivermectin_sensitive_subunits_from_both_C_elegans_and_H_contortus_rescue_the_drug_resistance_of_DA1316_/422933", "title"=>"Other ivermectin-sensitive subunits from both <i>C. elegans</i> and <i>H. contortus</i> rescue the drug resistance of DA1316.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 16:22:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/752679"], "description"=>"<p>The glutamate-gated chloride channel gene families of <i>C. elegans</i> and <i>H. contortus.</i></p>", "links"=>[], "tags"=>["glutamate-gated", "chloride"], "article_id"=>423052, "categories"=>["Genetics", "Infectious Diseases"], "users"=>["Susan K. Glendinning", "Steven D. Buckingham", "David B. Sattelle", "Susan Wonnacott", "Adrian J. Wolstenholme"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022390.t001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_glutamate_gated_chloride_channel_gene_families_of_C_elegans_and_H_contortus_/423052", "title"=>"The glutamate-gated chloride channel gene families of <i>C. elegans</i> and <i>H. contortus.</i>", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 16:23:30"}

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Relative Metric

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