title: Effect of the urease-derived peptide Jaburetox on the central nervous system of Triatoma infestans (Insecta: Heteroptera).
abstract: BACKGROUND
  Triatoma infestans is the main vector of Chagas'disease in Southern Cone countries. In triatomines, symptoms suggesting neurotoxicity were observed after treatment with Jaburetox (Jbtx), the entomotoxic peptide obtained from jackbean urease. Here, we study its effect in the central nervous system (CNS) of this species.
  
  
  METHODS
  Immunohistochemistry, Western blots, immunoprecipitation, two-dimensional electrophoresis, tandem mass spectrometry and enzymatic assays were performed.
  
  
  RESULTS
  Anti-Jbtx antibody labeled somata of the antennal lobe only in Jbtx-treated insects. Western blot assays of nervous tissue using the same antibody reacted with a 61kDa protein band only in peptide-injected insects. Combination of immunoprecipitation, two-dimensional electrophoresis and tandem mass spectrometry identified UDP-N-acetylglucosamine pyrophosphorylase (UDP-GlcNAcP) as a molecular target for Jbtx. The activity of UDP-GlcNAcP increased significantly in the CNS of Jbtx-treated insects. The effect of Jbtx on the activity of nitric oxide synthase (NOS) and NO production was investigated as NO is a recognized messenger molecule in the CNS of T. infestans. NOS activity and NO levels decreased significantly in CNS homogenates of Jbtx-treated insects.
  
  
  CONCLUSIONS
  UDP-GlcNAcP is a molecular target of Jbtx. Jbtx impaired the activity of T. infestans nitrergic system, which may be related with early behavioral effects.
  
  
  CONCLUSIONS
  We report that the CNS of Triatoma infestans is a target for the entomotoxic peptide and propose that a specific area of the brain is involved. Besides potentially providing tools for control strategies of Chagas' disease vectors our data may be relevant in various fields of research as insect physiology, neurobiology and protein function.
authors.affiliation: Facultad de Ciencias Biomédicas, Universidad Austral, Pilar, Provincia de Buenos Aires, Argentina; Museo Argentino de Ciencias Naturales, Ciudad Autónoma de Buenos Aires, Argentina. Electronic address: geronimogalvani@gmail.com.
authors.forename: Gerónimo L
authors.initials: GL
authors.lastname: Galvani
authors.affiliation: Dpto. Bioquímica Clínica, Centro de Investigaciones en Bioquímica Clínica e Inmunología (CIBICI-CONICET), Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Córdoba, Argentina. Electronic address: lfruttero@mail.fcq.unc.edu.ar.
authors.forename: Leonardo L
authors.initials: LL
authors.lastname: Fruttero
authors.affiliation: Laboratorio de Nocicepción y Dolor Neuropático, Instituto de Biología y Medicina Experimental - CONICET, Ciudad Autónoma de Buenos Aires, Argentina. Electronic address: mflorcoronel@yahoo.com.ar.
authors.forename: María F
authors.initials: MF
authors.lastname: Coronel
authors.affiliation: Centro de Investigaciones Endocrinólogicas "Dr. César Bergadá" (CEDIE-CONICET), Ciudad Autónoma de Buenos Aires, Argentina. Electronic address: snowicki@cedie.org.ar.
authors.forename: Susana
authors.initials: S
authors.lastname: Nowicki
authors.affiliation: Programa de Pós-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil. Electronic address: diogord@terra.com.br.
authors.forename: Diogo R
authors.initials: DR
authors.lastname: Demartini
authors.affiliation: Programa de Pós-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil. Electronic address: msdeff@gmail.com.
authors.forename: Marina S
authors.initials: MS
authors.lastname: Defferrari
authors.affiliation: Programa de Pós-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil. Electronic address: lisapostal@cbiot.ufrgs.br.
authors.forename: Melissa
authors.initials: M
authors.lastname: Postal
authors.affiliation: Dpto. Bioquímica Clínica, Centro de Investigaciones en Bioquímica Clínica e Inmunología (CIBICI-CONICET), Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Córdoba, Argentina. Electronic address: lcanavo@fcq.unc.edu.ar.
authors.forename: Lilián E
authors.initials: LE
authors.lastname: Canavoso
authors.affiliation: Programa de Pós-Graduação em Biologia Celular e Molecular, Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil; Departamento de Biofísica e Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil; Instituto do Cérebro-InsCer, Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre, RS, Brazil. Electronic address: celia.carlini@pucrs.br.
authors.forename: Célia R
authors.initials: CR
authors.lastname: Carlini
authors.affiliation: Facultad de Ciencias Biomédicas, Universidad Austral, Pilar, Provincia de Buenos Aires, Argentina; Museo Argentino de Ciencias Naturales, Ciudad Autónoma de Buenos Aires, Argentina. Electronic address: settembrini@macn.gov.ar.
authors.forename: Beatriz P
authors.initials: BP
authors.lastname: Settembrini
mesh_terms: D000818:Animals; D001522:Behavior, Animal / Q000187:drug effects; D002490:Central Nervous System / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000378:metabolism; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology*; D010940:Plant Proteins / Q000737:chemistry / Q000494:pharmacology*; D014224:Triatoma / Q000201:enzymology*; D014510:Urease / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D010455:Peptides; D010940:Plant Proteins; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D009713:Nucleotidyltransferases; C023226:UDPacetylglucosamine pyrophosphorylase; D014510:Urease
keywords: Canavalia ensiformis; Central nervous system; Nitric oxide; Triatoma infestans; UDP-N-acetylglucosaminepyrophosphorylase; Urease