title: Jack bean (Canavalia ensiformis) urease induces eicosanoid-modulated hemocyte aggregation in the Chagas' disease vector Rhodnius prolixus.
abstract: Ureases are multifunctional proteins that display biological activities independently of their enzymatic function, such as induction of exocytosis and insecticidal effects. Rhodnius prolixus, a major vector of Chagas' disease, is a model for studies on the entomotoxicity of jack bean urease (JBU). We have previously shown that JBU induces the production of eicosanoids in isolated tissues of R. prolixus. In insects, the immune response comprises cellular and humoral reactions, and is centrally modulated by eicosanoids. Cyclooxygenase products signal immunity in insects, mainly cellular reactions, such as hemocyte aggregation. In searching for a link between JBU's toxic effects and immune reactions in insects, we have studied the effects of this toxin on R. prolixus hemocytes. JBU triggers aggregation of hemocytes after injection into the hemocoel and when applied to isolated cells. On in vitro assays, the eicosanoid synthesis inhibitors dexamethasone (phospholipase A2 indirect inhibitor) and indomethacin (cyclooxygenase inhibitor) counteracted JBU's effect, indicating that eicosanoids, more specifically cyclooxygenase products, are likely to mediate the aggregation response. Contrarily, the inhibitors esculetin and baicalein were inactive, suggesting that lipoxygenase products are not involved in JBU's effect. Extracellular calcium was also necessary for JBU's effect, in agreement to other cell models responsive to ureases. A progressive darkening of the medium of JBU-treated hemocytes was observed, suggestive of a humoral response. JBU was immunolocalized in the cultured cells upon treatment along with cytoskeleton damage. The highest concentration of JBU tested on cultured cells also led to nuclei aggregation of adherent hemocytes. This is the first time urease has been shown to affect insect hemocytes, contributing to our understanding of the entomotoxic mechanisms of action of this protein.
authors.affiliation: Graduate Program in Cellular and Molecular Biology, Center of Biotechnology, Institute of Biosciences, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil. Electronic address: marina.defferrari@ufrgs.br.
authors.forename: M S
authors.initials: MS
authors.lastname: Defferrari
authors.affiliation: Department of Biology, McMaster University, Hamilton, ON, Canada. Electronic address: rosa.dasilva@mcmaster.ca.
authors.forename: R
authors.initials: R
authors.lastname: da Silva
authors.affiliation: Department of Biology, University of Toronto Mississauga, Mississauga, ON, Canada. Electronic address: ian.orchard@utoronto.ca.
authors.forename: I
authors.initials: I
authors.lastname: Orchard
authors.affiliation: Graduate Program in Cellular and Molecular Biology, Center of Biotechnology, Institute of Biosciences, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil; Instituto do Cérebro - InsCer, Pontificia Universidade Católica do Rio Grande do Sul, and Department of Biophysics, Biosciences Institute, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil. Electronic address: celia.carlini@pucrs.br.
authors.forename: C R
authors.initials: CR
authors.lastname: Carlini
mesh_terms: D000818:Animals; D001179:Arthropod Vectors / Q000502:physiology*; D032449:Canavalia / Q000737:chemistry* / Q000633:toxicity; D002449:Cell Aggregation / Q000187:drug effects; D014355:Chagas Disease / Q000635:transmission*; D015777:Eicosanoids / Q000096:biosynthesis / Q000633:toxicity*; D006434:Hemocytes / Q000187:drug effects*; D007814:Larva; D061251:Primary Cell Culture; D012239:Rhodnius / Q000502:physiology*; D014510:Urease / Q000633:toxicity*
chemical_list: D015777:Eicosanoids; D014510:Urease
keywords: Cyclooxygenase; Eicosanoids; Immune system; Immunocytochemistry; Insect; Toxicity