Functional and Genetic Characterization of Neuropeptide Y-Like Receptors in Aedes aegypti
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{"title"=>"Functional and Genetic Characterization of Neuropeptide Y-Like Receptors in Aedes aegypti", "type"=>"journal", "authors"=>[{"first_name"=>"Jeff", "last_name"=>"Liesch", "scopus_author_id"=>"55921611900"}, {"first_name"=>"Lindsay L.", "last_name"=>"Bellani", "scopus_author_id"=>"55922363400"}, {"first_name"=>"Leslie B.", "last_name"=>"Vosshall", "scopus_author_id"=>"6701388717"}], "year"=>2013, "source"=>"PLoS Neglected Tropical Diseases", "identifiers"=>{"sgr"=>"84887303297", "doi"=>"10.1371/journal.pntd.0002486", "pui"=>"370217996", "pmid"=>"24130914", "scopus"=>"2-s2.0-84887303297", "issn"=>"19352735", "isbn"=>"1935-2735 (Electronic)\\r1935-2727 (Linking)"}, "id"=>"cd48ac42-4582-300f-bc3c-db23c7dc1a0d", "abstract"=>"BACKGROUND: Female Aedes aegypti mosquitoes are the principal vector for dengue fever, causing 50-100 million infections per year, transmitted between human and mosquito by blood feeding. Ae. aegypti host-seeking behavior is known to be inhibited for three days following a blood meal by a hemolymph-borne humoral factor. Head Peptide-I is a candidate peptide mediating this suppression, but the mechanism by which this peptide alters mosquito behavior and the receptor through which it signals are unknown. METHODOLOGY/PRINCIPAL FINDINGS: Head Peptide-I shows sequence similarity to short Neuropeptide-F peptides (sNPFs) that have been implicated in feeding behaviors and are known to signal through Neuropeptide Y (NPY)-Like Receptors (NPYLRs). We identified eight NPYLRs in the Ae. aegypti genome and screened each in a cell-based calcium imaging assay for sensitivity against a panel of peptides. Four of the Ae. aegypti NPYLRs responded to one or more peptide ligands, but only NYPLR1 responded to Head Peptide-I as well as sNPFs. Two NPYLR1 homologues identified in the genome of the Lyme disease vector, Ixodes scapularis, were also sensitive to Head Peptide-I. Injection of synthetic Head Peptide-I and sNPF-3 inhibited host-seeking behavior in non-blood-fed female mosquitoes, whereas control injections of buffer or inactive Head Peptide-I [Cys10] had no effect. To ask if NPYLR1 is necessary for blood-feeding-induced host-seeking inhibition, we used zinc-finger nucleases to generate five independent npylr1 null mutant strains and tested them for behavioral abnormalities. npylr1 mutants displayed normal behavior in locomotion, egg laying, sugar feeding, blood feeding, host seeking, and inhibition of host seeking after a blood meal. CONCLUSIONS: In this work we deorphanized four Ae. aegypti NPYLRs and identified NPYLR1 as a candidate sNPF receptor that is also sensitive to Head Peptide-I. Yet npylr1 alone is not required for host-seeking inhibition and we conclude that other receptors, additional peptides, or both, regulate this important behavior.", "link"=>"http://www.mendeley.com/research/functional-genetic-characterization-neuropeptide-ylike-receptors-aedes-aegypti", "reader_count"=>58, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Librarian"=>2, "Researcher"=>13, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>4, "Student > Bachelor"=>6, "Lecturer"=>3, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Librarian"=>2, "Researcher"=>13, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>17, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>4, "Student > Bachelor"=>6, "Lecturer"=>3, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>38, "Medicine and Dentistry"=>5, "Arts and Humanities"=>1, "Neuroscience"=>3, "Philosophy"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Chemistry"=>1, "Social Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Neuroscience"=>{"Neuroscience"=>3}, "Chemistry"=>{"Chemistry"=>1}, "Social Sciences"=>{"Social Sciences"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>38}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>2}, "Arts and Humanities"=>{"Arts and Humanities"=>1}, "Philosophy"=>{"Philosophy"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "United Kingdom"=>1, "Portugal"=>1, "Indonesia"=>1}, "group_count"=>3}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1232872"], "description"=>"<p>(A) Diagram (left) and photograph (right) of the uniport olfactometer with a human hand presented as stimulus. (B) Attraction of female <i>Ae. aegypti</i> to a human host throughout the gonotrophic cycle (n = 3–5; ∼20 females per trial). Access to egg laying locations was permitted after 72 hours and observed at 96 hours. Data are plotted as mean±standard error of the mean (SEM). ANOVA with Dunnett's Correction for Multiple Comparison; *** = p<0.001, ns = not significant.</p>", "links"=>[], "tags"=>["mechanisms", "host-seeking", "inhibition"], "article_id"=>819960, "categories"=>["Information And Computing Sciences", "Biological Sciences", "Science Policy"], "users"=>["Jeff Liesch", "Lindsay L. Bellani", "Leslie B. Vosshall"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0002486.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Natural_mechanisms_for_female_host_seeking_inhibition_after_a_blood_meal_/819960", "title"=>"Natural mechanisms for female host-seeking inhibition after a blood meal.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-10 07:49:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1232874"], "description"=>"<p>(A) Phylogram depicting the genetic relationships of eight <i>Ae. aegypti</i> NPY-Like Receptors (NPYLR1, red; NPYLR2-8, black) as well as NPYLR1 homologues <i>An. gambiae</i>-sNPFR (blue), <i>Ix. scapularis</i> I.sca-NPYLR1A and I.sca-NPYLR1B (light and dark green), and <i>D. melanogaster</i>-sNPFR (magenta). (B) Peptides used in the HEK293T cell-based calcium imaging assay displayed with single letter amino acid codes and these variants: pE: pyroglutamic acid, hP: hydroxyproline, a: amidation, sNPF: <i>Ae. aegypti</i> short-Neuropeptide-F, NPF: <i>Ae. aegypti</i> Neuropeptide-F, Human NPY: Neuropeptide Y, Human PYY: Peptide YY. (C) Top: Representative images of HEK293T cells transiently transfected with <i>Ae. aegypti npylr1a</i>, illustrating calcium flux after application of indicated peptides. Bottom: Individual traces of fluorescent ratios representing Ca<sup>2+</sup> flux for the same HEK293T cells following application of the indicated peptides. (D) Head Peptide-I dose-response curves and EC50 values for NPYLR1 homologues in <i>Ix. scapularis</i>, <i>An. gambiae</i>, and <i>D. melanogaster</i>. (E) Summary of results for the cell-based calcium imaging screen. NPYLR2, NPYLR 3, NPYLR 4, and NPYLR 6 did not respond to any peptides tested (HP = Head Peptide).</p>", "links"=>[], "tags"=>["npy-like"], "article_id"=>819961, "categories"=>["Information And Computing Sciences", "Biological Sciences", "Science Policy"], "users"=>["Jeff Liesch", "Lindsay L. Bellani", "Leslie B. Vosshall"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0002486.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Deorphanization_of_NPY_Like_receptors_/819961", "title"=>"Deorphanization of NPY-Like receptors.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-10 07:49:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1232876"], "description"=>"<p>(A) Percent attraction of non-blood-fed female mosquitoes to human host stimuli after injection of buffer (open bar) or 4 and 10 mM of the indicated peptide. (n = 6; ∼20 females per trial, ANOVA with Bonferroni correction for multiple comparisons; ** = p<0.01). (B) qPCR analysis of <i>npylr1</i> transcripts in total RNA extracted from female whole body tissue at the indicated times after a blood meal. (n = 3, ANOVA-Tukey's Multiple Comparison of ΔCt values). All data are plotted as mean±SEM.</p>", "links"=>[], "tags"=>["peptide-i", "snpf-3", "inhibits", "host-seeking", "up-regulated"], "article_id"=>819963, "categories"=>["Information And Computing Sciences", "Biological Sciences", "Science Policy"], "users"=>["Jeff Liesch", "Lindsay L. Bellani", "Leslie B. Vosshall"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0002486.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Injection_of_Head_Peptide_I_or_sNPF_3_inhibits_host_seeking_behavior_and_npylr1_expression_is_up_regulated_after_blood_feeding_/819963", "title"=>"Injection of Head Peptide-I or sNPF-3 inhibits host-seeking behavior and <i>npylr1</i> expression is up-regulated after blood-feeding.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-10 07:49:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1232878"], "description"=>"<p>(A) Snake plot of predicted NPYLR1 membrane topology with the corresponding protein region encoded by the <i>npylr1</i> zinc-finger nuclease targeted DNA sequence colored in purple. (B) Illustration of the zinc-finger nuclease target region in the <i>npylr1</i> gene with corresponding mutations generated by non-homologous end-joining (<i>npylr1<sup>4</sup></i> and <i>npylr1<sup>8</sup></i>, red) and integration of a fluorescent marker by homologous recombination (<i>npylr1<sup>HR2</sup></i>, cyan). (C) Photographs of <i>npylr1<sup>HR2/+</sup></i> and wild-type mosquito pupae illustrating broad expression of the ECFP marker. (D) PCR verification of homologous recombination into the <i>npylr1</i> locus of the 2565 bp <i>polyubiquitin</i>>ECFP cassette. (E) Snake plots of predicted protein truncations for <i>npylr1</i> mutant alleles. Filled purple circles indicate ZFN target region and outlined purple circles indicate resulting frame-shift mutations.</p>", "links"=>[], "tags"=>["mutagenesis", "zinc-finger"], "article_id"=>819965, "categories"=>["Information And Computing Sciences", "Biological Sciences", "Science Policy"], "users"=>["Jeff Liesch", "Lindsay L. Bellani", "Leslie B. Vosshall"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0002486.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Targeted_mutagenesis_of_npylr1_using_zinc_finger_nucleases_/819965", "title"=>"Targeted mutagenesis of <i>npylr1</i> using zinc-finger nucleases.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-10 07:49:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1232879"], "description"=>"<p>(A) Average daily activity counts for individual female <i>npylr1<sup>HR2/HR3</sup></i> mutants (left, n = 14) and <i>npylr1<sup>8/4</sup></i> mutants (right, n = 7). (B) Eggs deposited per individual female <i>npylr1<sup>HR2/HR3</sup></i> (left, n = 18–20) and <i>npylr1<sup>8/4</sup></i> (right, n = 9–28). All data are plotted as mean±SEM. 1-Way ANOVA with Bonferroni correction for multiple comparisons. * = p<0.05; ** = p<0.01, ns = not significant.</p>", "links"=>[], "tags"=>["locomotor", "egg-laying"], "article_id"=>819966, "categories"=>["Information And Computing Sciences", "Biological Sciences", "Science Policy"], "users"=>["Jeff Liesch", "Lindsay L. Bellani", "Leslie B. Vosshall"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0002486.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Normal_locomotor_behavior_and_egg_laying_in_npylr1_mutants_/819966", "title"=>"Normal locomotor behavior and egg-laying in <i>npylr1</i> mutants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-10 07:49:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1232880"], "description"=>"<p>(A) Diagram of the CAFE assay. (B) Volume of sugar ingested for female mosquitoes in the CAFE after fasting for 0, 12, 24, 48, and 72 hours (n = 8–16, except 72 hours, n = 2; 5 mosquitoes per trial. Mean±SEM. 1-way ANOVA compared to 0 hours with Dunnett's multiple comparison test. * = p<0.05, *** = p<0.001, ns = not significant). (C) Sugar ingested by the indicated genotypes at 0 and 48 hours after fasting (n = 16; 5 females per trial. Mean±SEM, 2-way ANOVA with Bonferroni correction for multiple comparisons). (D) Diagram of the BUFFET assay. (E) Sugar ingested by the indicated genotypes at 48 hours after fasting (n = 64; 16 females tested in each of 4 independent trials). (F) Diagram of the membrane feeding assay. (G) Percent of mosquitoes for the indicated genotypes that blood-fed in the membrane feeding assay (n = 6, 15 females per trial). (H) The average blood meal sizes for the indicated genotypes. (n = 6, 1–12 females confirmed as fed per trial). Data in E, G, H are plotted as mean±SEM, 1-way ANOVA with Bonferroni correction for multiple comparisons.</p>", "links"=>[], "tags"=>["sugar-feeding", "blood-feeding"], "article_id"=>819967, "categories"=>["Information And Computing Sciences", "Biological Sciences", "Science Policy"], "users"=>["Jeff Liesch", "Lindsay L. Bellani", "Leslie B. Vosshall"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0002486.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Fasting_response_sugar_feeding_and_blood_feeding_are_normal_in_npylr1_mutants_/819967", "title"=>"Fasting response, sugar-feeding and blood-feeding are normal in <i>npylr1</i> mutants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-10 07:49:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1232881"], "description"=>"<p>Host-seeking behavior of <i>npylr1</i> (A) homologous recombination mutants and (B) non-homologous end-joining mutants with associated controls before and 24, 48, 72, and 120 hours after a blood meal. Egg-laying was observed after 72 hours and completed by 120 hours (n = 6–12, ∼20 females per trial). (C) Host-seeking behavior of <i>npylr1</i> homologous recombination mutants and associated controls at 32 and 40 hours after a blood meal (n = 3, ∼20 females per trial). All data are plotted as mean±SEM. 1-way ANOVA with Bonferroni correction for multiple comparisons within each time point.</p>", "links"=>[], "tags"=>["host-seeking", "mutants", "gonotrophic"], "article_id"=>819968, "categories"=>["Information And Computing Sciences", "Biological Sciences", "Science Policy"], "users"=>["Jeff Liesch", "Lindsay L. Bellani", "Leslie B. Vosshall"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0002486.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Normal_host_seeking_behavior_of_npylr1_mutants_throughout_the_gonotrophic_cycle_/819968", "title"=>"Normal host-seeking behavior of <i>npylr1</i> mutants throughout the gonotrophic cycle.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-10 07:49:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1232882"], "description"=>"<p>(A) Model 1 illustrates the hypothesis that known peptides including Head Peptide-I and sNPFs signal through an unknown receptor (“Receptor X”) and that NPYLR1 is not involved in the inhibition of host-seeking behavior. (B) Model 2 illustrates the hypothesis that a combinatorial network of redundant mechanisms for behavioral inhibition may exist after blood-feeding.</p>", "links"=>[], "tags"=>["inhibition", "host-seeking"], "article_id"=>819969, "categories"=>["Information And Computing Sciences", "Biological Sciences", "Science Policy"], "users"=>["Jeff Liesch", "Lindsay L. Bellani", "Leslie B. Vosshall"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0002486.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Models_for_the_inhibition_of_host_seeking_behavior_in_Ae_aegypti_/819969", "title"=>"Models for the inhibition of host-seeking behavior in <i>Ae. aegypti</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-10 07:49:54"}

PMC Usage Stats | Further Information

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Relative Metric

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